Compositions and Methods for Antibodies Targeting Complement Protein C5

ABSTRACT

The present invention relates to antibodies targeting complement protein C5 and compositions and methods of use thereof.

This application claims priority to U.S. Provisional Application Ser.No. 61/086,355 filed 5 Aug. 2008, the contents of which are incorporatedherein by reference in their entirety.

1. INTRODUCTION

The present invention relates to antibodies targeting complement proteinC5 and compositions and methods of use thereof.

2. BACKGROUND OF THE INVENTION

The normal role of complement, which is part of the innate immunesystem, is in host defense. Complement defends against bacterialinfection, links adaptive and innate immunity, and disposes immunecomplexes and the products of inflammatory injury.

The defensive functions are accomplished by biologically active productsgenerated in the course of complement activation, which opsoniseinfectious agents, promote inflammation or lyse susceptible targets(Marzari et al., Eur J Immunol 32:2773-2782 (2002)). The complementsystem consists of about 25-30 plasma proteins which play a role in theimmune system. The complement cascade is activated by at least threemajor pathways. The classical pathway is typically activated byimmune-complexes, the alternative pathway can be activated byunprotected cell surfaces, and the mannose binding lectin (MBL) pathwayis initiated by binding of MBL to cell surface carbohydrates(Trendelenburg, Swiss Med Wkly 137:413-417 (2007)).

All three pathways lead to the cleavage of C5 by the C5 convertase. Theresult of this cleavage is release of C5a fragment, a potentinflammatory molecule, and C5b which initiates the membrane attackcomplex (MAC). The complement products, once released, do notdifferentiate between foreign and self targets and, if not tightlyregulated, often cause extensive damage of bystander cells and tissuesin clinical conditions associated with unrestricted complementactivation (Marzari et al., 2002).

C5 is expressed intracellularly as a single pro-C5 peptide of 1676 aminoacids that consist of an 18 residue signal sequence and an Arg-richlinker sequence (RPRR) situated between the mature N-terminal β chainand the C-terminal α chain. The mature C5 has a molecular weight ofabout 190 kDa, and consists of two polypeptide chains (α, 115 kDa and β,75 kDa) which are connected by disulfide bonds. The C5 convertasecleaves C5 between residues 74 and 75 of the alpha chain to release the74 amino acid C5a peptide and the C5b fragment which is subsequentlyincorporated into the membrane-attack complex (MAC).

Macular degeneration is a medical condition predominantly found in theelderly in which the center of the inner lining of the eye, known as themacula area of the retina, suffers thinning, atrophy, and in some cases,bleeding. This can result in loss of central vision, which entailsinability to see fine details, to read, or to recognize faces.Pathogenesis of new choroidal vessel formation is poorly understood, butfactors such as inflammation, ischemia, and local production ofangiogenic factors are thought to be important.

Despite current treatment options for treating diseases and disordersassociated with the classical or alternative component pathways,particularly AMD, there remains a need for finding specific targets thatlead to treatments which are effective and well-tolerated.

3. SUMMARY OF THE INVENTION

The present invention provides isolated complement C5-binding molecules(e.g., C5-binding antibodies or antigen binding fragments thereof),pharmaceutical compositions comprising such molecules, methods of makingsuch molecules and compositions, and methods of use thereof.

In some embodiments, the present invention provides isolated antibodiesor antigen binding fragments thereof that specifically bind to a C5protein, wherein said antibody has an affinity constant (K_(A)) of atleast 1×10⁷ M⁻¹, 10⁸ M⁻¹, 10⁹ M⁻¹, 10¹⁰ M⁻¹, or 10¹¹ M⁻¹.

In some embodiments, the present invention provides isolated antibodiesor antigen binding fragments thereof that specifically bind to a C5protein, and inhibit the alternative complete pathway as measured by invitro hemolytic assay with an IC₅₀ range from about 20 pM to about 200pM.

In some embodiments, the present invention provides isolated antibodiesor antigen binding fragments thereof that specifically bind to a C5protein, and cross compete with an antibody described in Table 1 below.In some embodiments, the present invention provides isolated antibodiesor antigen binding fragments thereof that bind to the same epitope of C5protein as an antibody described in Table 1 below.

In some embodiments, the antibodies of the invention are isolatedmonoclonal antibodies that specifically bind to a C5 protein. In someembodiments, the antibodies of the invention are isolated human orhumanized monoclonal antibodies that specifically bind to a C5 protein.In some embodiments, the antibodies of the invention are isolatedchimeric antibodies that specifically bind to a C5 protein. In someembodiments, the antibodies of the invention comprise a human heavychain constant region and a human light chain constant region.

In some embodiments, the present invention provides isolated antibodiesor antigen binding fragments thereof that specifically bind to a C5protein, wherein said antibodies are single chain antibodies. In someembodiments, the antibodies of the invention are Fab fragments. In someembodiments, the antibodies of the invention are scFv.

In some embodiments, the present invention provides isolated antibodiesor antigen-binding fragments thereof that specifically bind to bothhuman C5 and cynomolgus C5. In some embodiments, the antibodies of theinvention are an IgG isotype.

In some embodiments, the present invention provides isolated antibodiesor antigen binding fragments thereof comprising a framework in whichamino acids have been substituted into the antibody framework from therespective human VH or VL germline sequences.

In some embodiments, the present invention provides isolated monoclonalantibodies or antigen binding fragments thereof that specifically bindto a C5 protein, wherein said antibodies comprise at least onecomplementarity determining (CDR) sequence having at least 90%, 95%,97%, 98% or at least 99% sequence identity to SEQ ID NO: 1, 2, 3, 4, 5,6, 17, 18, 19, 20, 21, 22, 33, 34, 35, 36, 37, 38, 49, 50, 61, 62, 63,64, 65, 66, 77, 78, 89, 95, 101, 107, 113, 119, 120, 131, 132, 133, 134,135, 136, 145, 146, 147, 148, 149, 150, 159, 160, 161, 162, 163, 164,173, 174, 175, 176, 177, 178, 195, 196, 197, 198, 199, 200, 209, 226,235, 236, 237, 238, 239, or 240.

In some embodiments, the present invention provides isolated monoclonalantibodies or antigen binding fragments thereof that specifically bindto a C5 protein, wherein said antibodies comprise at least one heavychain CDR sequence that is identical to SEQ ID NO: 1, 2, 3, 17, 18, 19,33, 34, 35, 49, 61, 62, 63, 77, 77, 95, 107, 113, 119, 132, 131, 133,145, 146, 147, 159, 160, 161, 173, 174, 175, 195, 196, 197, 226, 235,236, or 237.

In some embodiments, the present invention provides isolated monoclonalantibodies or antigen binding fragments thereof that specifically bindto a C5 protein, wherein said antibodies comprise at least one lightchain CDR sequence that is identical to SEQ ID NO: 4, 5, 6, 20, 21, 22,36, 37, 38, 50, 64, 65, 66, 78, 89, 101, 120, 134, 135, 136, 148, 149,150, 162, 163, 164, 176, 177, 178, 198, 199, 200, 209, 238, 239, or 240.

In some embodiments, the present invention provides isolated monoclonalantibodies or antigen binding fragments thereof that specifically bindto a C5 protein, wherein said antibodies comprise a heavy chain CDR 1selected from the group consisting SEQ ID NOs: 1, 17, 33, 61, 131, 145,159, 173, 195, and 235; a heavy chain CDR2 selected from the groupconsisting SEQ ID NOs: 2, 18, 34, 49, 62, 77, 95, 107, 113, 119, 132,146, 160, 174, 196, 226, and 236; and a heavy chain CDR3 selected fromthe group consisting SEQ ID NOs: 3, 19, 35, 63, 133, 147, 161, 175, 197,and 237. In some embodiments, such antibodies or antigen bindingfragments thereof further comprise a light chain CDR1 selected from thegroup consisting SEQ ID NOs: 4, 20, 36, 64, 134, 148, 162, 176, 198, and238; a light chain CDR2 selected from the group consisting SEQ ID NOs:5, 21, 37, 65, 135, 149, 163, 177, 199, and 239; and a light chain CDR3selected from the group consisting SEQ ID NOs: 6, 22, 38, 50, 66, 78,89, 101, 120, 136, 150, 164, 178, 200, 209, and 240.

In some embodiments, the present invention provides isolated monoclonalantibodies or antigen binding fragments thereof that specifically bindto a C5 protein, wherein said antibodies comprise a light chain CDR 1selected from the group consisting SEQ ID NOs: 4, 20, 36, 64, 134, 148,162, 176, 198, and 238; a light chain CDR2 selected from the groupconsisting SEQ ID NOs: 5, 21, 37, 65, 135, 149, 163, 177, 199, and 239;and a light chain CDR3 selected from the group consisting SEQ ID NOs: 6,22, 38, 50, 66, 78, 89, 101, 120, 136, 150, 164, 178, 200, 209, and 240.

In some embodiments, the present invention provides isolated monoclonalantibodies or antigen binding fragments thereof that specifically bindto a C5 protein, wherein said antibodies comprise a heavy chain variableregion having at least 90%, 95%, 97%, 98% or at least 99% sequenceidentity to SEQ ID NO: 7, 23, 39, 51, 67, 79, 96, 108, 114, 121, 137,151, 165, 179, 187, 201, 210, 218, 227, 241, 253, 257, 273, 277, or 281.In some embodiments, such antibodies or antigen binding fragmentsthereof further comprise a light chain variable region having at least90%, 95%, 97%, 98% or at least 99% sequence identity to SEQ ID NO: 8,24, 40, 52, 68, 80, 90, 102, 122, 138, 152, 166, 180, 188, 202, 211,219, 228, 242, 261, 265, 269, 285, and 289.

In some embodiments, the present invention provides isolated monoclonalantibodies or antigen binding fragments thereof that specifically bindto a C5 protein, wherein said antibodies comprise a light chain variableregion having at least 90%, 95%, 97%, 98% or at least 99% sequenceidentity to SEQ ID NO: 8, 24, 40, 52, 68, 80, 90, 102, 122, 138, 152,166, 180, 188, 202, 211, 219, 228, 242, 261, 265, 269, 285, and 289.

In some embodiments, the present invention provides isolated monoclonalantibodies or antigen binding fragments thereof that specifically bindto a C5 protein, wherein said antibodies comprise a heavy chain havingat least 90%, 95%, 97%, 98% or at least 99% sequence identity to SEQ IDNO: 9, 25, 41, 53, 69, 81, 97, 109, 115, 123, 139, 153, 167, 181, 189,203, 212, 220, 229, 243, 249, 254, 258, 274, 278, or 282. In someembodiments, such antibodies further comprise a light chain having atleast 90%, 95%, 97%, 98% or at least 99% sequence identity to SEQ ID NO:10, 26, 42, 54, 70, 82, 91, 103, 124, 140, 154, 168, 182, 190, 204, 213,221, 230, 244, 251, 262, 266, 270, 286, or 290.

In some embodiments, the present invention provides isolated monoclonalantibodies or antigen binding fragments thereof that specifically bindto a C5 protein, wherein said antibodies comprise a light chain havingat least 90%, 95%, 97%, 98% or at least 99% sequence identity to SEQ IDNO: 10, 26, 42, 54, 70, 82, 91, 103, 124, 140, 154, 168, 182, 190, 204,213, 221, 230, 244, 251, 262, 266, 270, 286, or 290.

The present invention also comprises pharmaceutical compositionscomprising one or more C5-binding molecules of the invention (e.g., C5binding antibodies or antigen binding fragments thereof) and apharmaceutically acceptable carrier.

In some embodiments, the present invention provides nucleic acidscomprising a nucleotide sequence encoding a polypeptide comprising aheavy chain variable region having at least 90%, 95%, 97%, 98% or atleast 99% sequence identity to SEQ ID NO: 7, 23, 39, 51, 67, 79, 96,108, 114, 121, 137, 151, 165, 179, 187, 201, 210, 218, 227, 241, 253,257, 273, 277, or 281.

In some embodiments, the present invention provides nucleic acidscomprising a nucleotide sequence encoding a polypeptide comprising alight chain variable region having at least 90%, 95%, 97%, 98% or atleast 99% sequence identity to SEQ ID NO: 8, 24, 40, 52, 68, 80, 90,102, 122, 138, 152, 166, 180, 188, 202, 211, 219, 228, 242, 261, 265,269, 285, and 289.

The present invention also provides vectors and host cells comprisingsuch nucleic acids. In one embodiment, the present invention providesisolated host cells comprising (1) a recombinant DNA segment encoding aheavy chain of the antibodies of the invention, and (2) a secondrecombinant DNA segment encoding a light chain of the antibodies of theinvention; wherein said DNA segments are respectively operably linked toa first and a second promoter, and are capable of being expressed insaid host cell. In another embodiment, the present invention providesisolated host cells comprising a recombinant DNA segment encoding aheavy chain, and a light chain of the antibodies of the invention,respectively, wherein said DNA segment is operably linked to a promoter,and is capable of being expressed in said host cells. In someembodiments, the host cells are non-human mammalian cell line. In someembodiments, the antibodies or antigen binding fragments thereof are ahuman monoclonal antibody, or an antigen binding fragment thereof.

The present invention further provides treatment of diagnostic methodsusing the C5 binding molecules (e.g., C5 binding antibodies or antigenbinding fragments thereof) of the invention. In one embodiment, thepresent invention provides methods of treating age related maculardegeneration comprising administering to a subject in need thereof aneffective amount of a composition comprising an antibody or an antigenbinding fragment thereof of the invention.

In another embodiment, the present invention provides methods oftreating a disease comprising administering to a subject in need thereofan effective amount of a composition comprising an antibody or anantigen binding fragment thereof of the invention, wherein said diseaseis asthma, arthritis, autoimmune heart disease, multiple sclerosis,inflammatory bowel disease, ischemia-reperfusion injuries,Barraquer-Simons Syndrome, hemodialysis, systemic lupus, lupuserythematosus, psoriasis, multiple sclerosis, transplantation,Alzheimer's disease, glomerulonephritis, or MPGN II.

The present invention also provides methods of treating paroxysmalnocturnal hemoglobinuria (PNH) comprising administering to a subject inneed thereof an effective amount of a composition comprising an antibodyor antigen binding fragment thereof of the invention.

The present invention further provides methods of ameliorating a symptomassociated with extracorporeal circulation comprising administering to asubject in need thereof an effective amount of a composition comprisingan antibody or antigen binding fragment thereof of the invention.

3.1. DEFINITIONS

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by those of ordinary skillin the art to which this invention pertains.

The term “antibody” as used herein includes whole antibodies and anyantigen binding fragment (i.e., “antigen-binding portion”) or singlechains thereof. A naturally occurring “antibody” is a glycoproteincomprising at least two heavy (H) chains and two light (L) chainsinter-connected by disulfide bonds. Each heavy chain is comprised of aheavy chain variable region (abbreviated herein as VH) and a heavy chainconstant region. The heavy chain constant region is comprised of threedomains, CH1, CH2 and CH3. Each light chain is comprised of a lightchain variable region (abbreviated herein as VL) and a light chainconstant region. The light chain constant region is comprised of onedomain, CL. The VH and VL regions can be further subdivided into regionsof hypervariability, termed complementarity determining regions (CDR),interspersed with regions that are more conserved, termed frameworkregions (FR). Each VH and VL is composed of three CDRs and four FRsarranged from amino-terminus to carboxy-terminus in the following order:FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavyand light chains contain a binding domain that interacts with anantigen. The constant regions of the antibodies may mediate the bindingof the immunoglobulin to host tissues or factors, including variouscells of the immune system (e.g., effector cells) and the firstcomponent (C1q) of the classical complement system.

The term “antigen binding portion” of an antibody, as used herein,refers to one or more fragments of an intact antibody that retain theability to specifically bind to a given antigen (e.g., C5). Antigenbinding functions of an antibody can be performed by fragments of anintact antibody. Examples of binding fragments encompassed within theterm “antigen binding portion” of an antibody include a Fab fragment, amonovalent fragment consisting of the VL, VH, CL and CH1 domains; aF(ab)₂ fragment, a bivalent fragment comprising two Fab fragments linkedby a disulfide bridge at the hinge region; an Fd fragment consisting ofthe VH and CH1 domains; an Fv fragment consisting of the VL and VHdomains of a single arm of an antibody; a single domain antibody (dAb)fragment (Ward et al., 1989 Nature 341:544-546), which consists of a VHdomain; and an isolated complementarity determining region (CDR).

Furthermore, although the two domains of the Fv fragment, VL and VH, arecoded for by separate genes, they can be joined, using recombinantmethods, by an artificial peptide linker that enables them to be made asa single protein chain in which the VL and VH regions pair to formmonovalent molecules (known as single chain Fv (scFv); see, e.g., Birdet al., 1988 Science 242:423-426; and Huston et al., 1988 Proc. Natl.Acad. Sci. 85:5879-5883). Such single chain antibodies include one ormore “antigen binding portions” of an antibody. These antibody fragmentsare obtained using conventional techniques known to those of skill inthe art, and the fragments are screened for utility in the same manneras are intact antibodies.

Antigen binding portions can also be incorporated into single domainantibodies, maxibodies, minibodies, intrabodies, diabodies, triabodies,tetrabodies, v-NAR and bis-scFv (see, e.g., Hollinger and Hudson, 2005,Nature Biotechnology, 23, 9, 1126-1136). Antigen binding portions ofantibodies can be grafted into scaffolds based on polypeptides such asFibronectin type III (Fn3) (see U.S. Pat. No. 6,703,199, which describesfibronectin polypeptide monobodies).

Antigen binding portions can be incorporated into single chain moleculescomprising a pair of tandem Fv segments (VH-CH1-VH-CH1) which, togetherwith complementary light chain polypeptides, form a pair of antigenbinding regions (Zapata et al., 1995 Protein Eng. 8(10):1057-1062; andU.S. Pat. No. 5,641,870).

As used herein, the term “Affinity” refers to the strength ofinteraction between antibody and antigen at single antigenic sites.Within each antigenic site, the variable region of the antibody “arm”interacts through weak non-covalent forces with antigen at numeroussites; the more interactions, the stronger the affinity.

As used herein, the term “Avidity” refers to an informative measure ofthe overall stability or strength of the antibody-antigen complex. It iscontrolled by three major factors: antibody epitope affinity; thevalency of both the antigen and antibody; and the structural arrangementof the interacting parts. Ultimately these factors define thespecificity of the antibody, that is, the likelihood that the particularantibody is binding to a precise antigen epitope.

The term “amino acid” refers to naturally occurring and synthetic aminoacids, as well as amino acid analogs and amino acid mimetics thatfunction in a manner similar to the naturally occurring amino acids.Naturally occurring amino acids are those encoded by the genetic code,as well as those amino acids that are later modified, e.g.,hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. Amino acidanalogs refer to compounds that have the same basic chemical structureas a naturally occurring amino acid, i.e., an alpha carbon that is boundto a hydrogen, a carboxyl group, an amino group, and an R group, e.g.,homoserine, norleucine, methionine sulfoxide, methionine methylsulfonium. Such analogs have modified R groups (e.g., norleucine) ormodified peptide backbones, but retain the same basic chemical structureas a naturally occurring amino acid. Amino acid mimetics refers tochemical compounds that have a structure that is different from thegeneral chemical structure of an amino acid, but that functions in amanner similar to a naturally occurring amino acid.

The term “binding specificity” as used herein refers to the ability ofan individual antibody combining site to react with only one antigenicdeterminant. The combining site of the antibody is located in the Fabportion of the molecule and is constructed from the hypervariableregions of the heavy and light chains. Binding affinity of an antibodyis the strength of the reaction between a single antigenic determinantand a single combining site on the antibody. It is the sum of theattractive and repulsive forces operating between the antigenicdeterminant and the combining site of the antibody.

Specific binding between two entities means a binding with anequilibrium constant (K_(A)) of at least 1×10⁷ M⁻¹, 10⁸ M⁻¹, 10⁹ M⁻¹,10¹⁰ M⁻¹, or 10¹¹ M⁻¹. The phrase “specifically (or selectively) binds”to an antibody (e.g., a C5-binding antibody) refers to a bindingreaction that is determinative of the presence of a cognate antigen(e.g., a human C5 or cynomolgus C5) in a heterogeneous population ofproteins and other biologics. In addition to the equilibrium constant(KA) noted above, an C5-binding antibody of the invention typically alsohas a dissociation rate constant (Kd) of about 1×10⁻² s⁻¹, 1×10⁻³ s⁻¹,1×10⁻⁴ s⁻¹, 1×10⁻⁴ s⁻¹, or lower, and binds to C5 with an affinity thatis at least two-fold greater than its affinity for binding to anon-specific antigen (e.g., C3, C4, BSA). The phrases “an antibodyrecognizing an antigen” and “an antibody specific for an antigen” areused interchangeably herein with the term “an antibody which bindsspecifically to an antigen”.

The term “chimeric antibody” is an antibody molecule in which (a) theconstant region, or a portion thereof, is altered, replaced or exchangedso that the antigen binding site (variable region) is linked to aconstant region of a different or altered class, effector functionand/or species, or an entirely different molecule which confers newproperties to the chimeric antibody, e.g., an enzyme, toxin, hormone,growth factor, drug, etc.; or (b) the variable region, or a portionthereof, is altered, replaced or exchanged with a variable region havinga different or altered antigen specificity. For example, a mouseantibody can be modified by replacing its constant region with theconstant region from a human immunoglobulin. Due to the replacement witha human constant region, the chimeric antibody can retain itsspecificity in recognizing the antigen while having reduced antigenicityin human as compared to the original mouse antibody.

The term “complement C5 protein” or “C5” are used interchangeably, andrefers to the C5 protein in different species. For example, human C5 hasthe sequence as set in SEQ ID NO: 296, cynomolgus C5 has the sequence asset in SEQ ID NO: 297 (Macaca fascicularis) (see Table 1). Human C5 canbe obtained from Quidel (Cat. Number A403). Cynomolgus C5 can beproduced as illustrated in the Example section below.

The term “conservatively modified variant” applies to both amino acidand nucleic acid sequences. With respect to particular nucleic acidsequences, conservatively modified variants refers to those nucleicacids which encode identical or essentially identical amino acidsequences, or where the nucleic acid does not encode an amino acidsequence, to essentially identical sequences. Because of the degeneracyof the genetic code, a large number of functionally identical nucleicacids encode any given protein. For instance, the codons GCA, GCC, GCGand GCU all encode the amino acid alanine. Thus, at every position wherean alanine is specified by a codon, the codon can be altered to any ofthe corresponding codons described without altering the encodedpolypeptide. Such nucleic acid variations are “silent variations,” whichare one species of conservatively modified variations. Every nucleicacid sequence herein which encodes a polypeptide also describes everypossible silent variation of the nucleic acid. One of skill willrecognize that each codon in a nucleic acid (except AUG, which isordinarily the only codon for methionine, and TGG, which is ordinarilythe only codon for tryptophan) can be modified to yield a functionallyidentical molecule. Accordingly, each silent variation of a nucleic acidthat encodes a polypeptide is implicit in each described sequence.

For polypeptide sequences, “conservatively modified variants” includeindividual substitutions, deletions or additions to a polypeptidesequence which result in the substitution of an amino acid with achemically similar amino acid. Conservative substitution tablesproviding functionally similar amino acids are well known in the art.Such conservatively modified variants are in addition to and do notexclude polymorphic variants, interspecies homologs, and alleles of theinvention. The following eight groups contain amino acids that areconservative substitutions for one another: 1) Alanine (A), Glycine (G);2) Aspartic acid (D), Glutamic acid (E); 3) Asparagine (N), Glutamine(Q); 4) Arginine (R), Lysine (K); 5) Isoleucine (I), Leucine (L),Methionine (M), Valine (V); 6) Phenylalanine (F), Tyrosine (Y),Tryptophan (W); 7) Serine (S), Threonine (T); and 8) Cysteine (C),Methionine (M) (see, e.g., Creighton, Proteins (1984)). In someembodiments, the term “conservative sequence modifications” are used torefer to amino acid modifications that do not significantly affect oralter the binding characteristics of the antibody containing the aminoacid sequence.

The terms “cross-block”, “cross-blocked” and “cross-blocking” are usedinterchangeably herein to mean the ability of an antibody or otherbinding agent to interfere with the binding of other antibodies orbinding agents to C5 in a standard competitive binding assay.

The ability or extent to which an antibody or other binding agent isable to interfere with the binding of another antibody or bindingmolecule to C5, and therefore whether it can be said to cross-blockaccording to the invention, can be determined using standard competitionbinding assays. One suitable assay involves the use of the Biacoretechnology (e.g. by using the BIAcore 3000 instrument (Biacore, Uppsala,Sweden)), which can measure the extent of interactions using surfaceplasmon resonance technology. Another assay for measuring cross-blockinguses an ELISA-based approach.

The term “epitope” means a protein determinant capable of specificbinding to an antibody. Epitopes usually consist of chemically activesurface groupings of molecules such as amino acids or sugar side chainsand usually have specific three dimensional structural characteristics,as well as specific charge characteristics. Conformational andnonconformational epitopes are distinguished in that the binding to theformer but not the latter is lost in the presence of denaturingsolvents.

As used herein, the term “high affinity” for an IgG antibody refers toan antibody having a KD of 10⁻⁸ M or less, 10⁻⁹ M or less, or 10⁻¹⁰ M,or 10⁻¹¹ M or less for a target antigen. However, “high affinity”binding can vary for other antibody isotypes. For example, “highaffinity” binding for an IgM isotype refers to an antibody having a KDof 10⁻⁷ M or less, or 10⁻⁸ M or less.

The term “human antibody”, as used herein, is intended to includeantibodies having variable regions in which both the framework and CDRregions are derived from sequences of human origin. Furthermore, if theantibody contains a constant region, the constant region also is derivedfrom such human sequences, e.g., human germline sequences, or mutatedversions of human germline sequences. The human antibodies of theinvention may include amino acid residues not encoded by human sequences(e.g., mutations introduced by random or site-specific mutagenesis invitro or by somatic mutation in vivo).

The term “human monoclonal antibody” refers to antibodies displaying asingle binding specificity which have variable regions in which both theframework and CDR regions are derived from human sequences. In oneembodiment, the human monoclonal antibodies are produced by a hybridomawhich includes a B cell obtained from a transgenic nonhuman animal,e.g., a transgenic mouse, having a genome comprising a human heavy chaintransgene and a light chain transgene fused to an immortalized cell.

A “humanized” antibody is an antibody that retains the reactivity of anon-human antibody while being less immunogenic in humans. This can beachieved, for instance, by retaining the non-human CDR regions andreplacing the remaining parts of the antibody with their humancounterparts (i.e., the constant region as well as the frameworkportions of the variable region). See, e.g., Morrison et al., Proc.Natl. Acad. Sci. USA, 81:6851-6855, 1984; Morrison and Oi, Adv.Immunol., 44:65-92, 1988; Verhoeyen et al., Science, 239:1534-1536,1988; Padlan, Molec. Immun., 28:489-498, 1991; and Padlan, Molec.Immun., 31:169-217, 1994. Other examples of human engineering technologyinclude, but is not limited to Xoma technology disclosed in U.S. Pat.No. 5,766,886.

The terms “identical” or percent “identity,” in the context of two ormore nucleic acids or polypeptide sequences, refer to two or moresequences or subsequences that are the same. Two sequences are“substantially identical” if two sequences have a specified percentageof amino acid residues or nucleotides that are the same (i.e., 60%identity, optionally 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identityover a specified region, or, when not specified, over the entiresequence), when compared and aligned for maximum correspondence over acomparison window, or designated region as measured using one of thefollowing sequence comparison algorithms or by manual alignment andvisual inspection. Optionally, the identity exists over a region that isat least about 50 nucleotides (or 10 amino acids) in length, or morepreferably over a region that is 100 to 500 or 1000 or more nucleotides(or 20, 50, 200 or more amino acids) in length.

For sequence comparison, typically one sequence acts as a referencesequence, to which test sequences are compared. When using a sequencecomparison algorithm, test and reference sequences are entered into acomputer, subsequence coordinates are designated, if necessary, andsequence algorithm program parameters are designated. Default programparameters can be used, or alternative parameters can be designated. Thesequence comparison algorithm then calculates the percent sequenceidentities for the test sequences relative to the reference sequence,based on the program parameters.

A “comparison window”, as used herein, includes reference to a segmentof any one of the number of contiguous positions selected from the groupconsisting of from 20 to 600, usually about 50 to about 200, moreusually about 100 to about 150 in which a sequence may be compared to areference sequence of the same number of contiguous positions after thetwo sequences are optimally aligned. Methods of alignment of sequencesfor comparison are well known in the art. Optimal alignment of sequencesfor comparison can be conducted, e.g., by the local homology algorithmof Smith and Waterman (1970) Adv. Appl. Math. 2:482c, by the homologyalignment algorithm of Needleman and Wunsch, J. Mol. Biol. 48:443, 1970,by the search for similarity method of Pearson and Lipman, Proc. Nat'l.Acad. Sci. USA 85:2444, 1988, by computerized implementations of thesealgorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin GeneticsSoftware Package, Genetics Computer Group, 575 Science Dr., Madison,Wis.), or by manual alignment and visual inspection (see, e.g., Brent etal., Current Protocols in Molecular Biology, John Wiley & Sons, Inc.(ringbou ed., 2003)).

Two examples of algorithms that are suitable for determining percentsequence identity and sequence similarity are the BLAST and BLAST 2.0algorithms, which are described in Altschul et al., Nuc. Acids Res.25:3389-3402, 1977; and Altschul et al., J. Mol. Biol. 215:403-410,1990, respectively. Software for performing BLAST analyses is publiclyavailable through the National Center for Biotechnology Information.This algorithm involves first identifying high scoring sequence pairs(HSPs) by identifying short words of length W in the query sequence,which either match or satisfy some positive-valued threshold score Twhen aligned with a word of the same length in a database sequence. T isreferred to as the neighborhood word score threshold (Altschul et al.,supra). These initial neighborhood word hits act as seeds for initiatingsearches to find longer HSPs containing them. The word hits are extendedin both directions along each sequence for as far as the cumulativealignment score can be increased. Cumulative scores are calculatedusing, for nucleotide sequences, the parameters M (reward score for apair of matching residues; always >0) and N (penalty score formismatching residues; always <0). For amino acid sequences, a scoringmatrix is used to calculate the cumulative score. Extension of the wordhits in each direction are halted when: the cumulative alignment scorefalls off by the quantity X from its maximum achieved value; thecumulative score goes to zero or below, due to the accumulation of oneor more negative-scoring residue alignments; or the end of eithersequence is reached. The BLAST algorithm parameters W, T, and Xdetermine the sensitivity and speed of the alignment. The BLASTN program(for nucleotide sequences) uses as defaults a wordlength (N) of 11, anexpectation (E) or 10, M=5, N=−4 and a comparison of both strands. Foramino acid sequences, the BLASTP program uses as defaults a wordlengthof 3, and expectation (E) of 10, and the BLOSUM62 scoring matrix (seeHenikoff and Henikoff, Proc. Natl. Acad. Sci. USA 89:10915, 1989)alignments (B) of 50, expectation (E) of 10, M=5, N=−4, and a comparisonof both strands.

The BLAST algorithm also performs a statistical analysis of thesimilarity between two sequences (see, e.g., Karlin and Altschul, Proc.Natl. Acad. Sci. USA 90:5873-5787, 1993). One measure of similarityprovided by the BLAST algorithm is the smallest sum probability (P(N)),which provides an indication of the probability by which a match betweentwo nucleotide or amino acid sequences would occur by chance. Forexample, a nucleic acid is considered similar to a reference sequence ifthe smallest sum probability in a comparison of the test nucleic acid tothe reference nucleic acid is less than about 0.2, more preferably lessthan about 0.01, and most preferably less than about 0.001.

The percent identity between two amino acid sequences can also bedetermined using the algorithm of E. Meyers and W. Miller (Comput. Appl.Biosci., 4:11-17, 1988) which has been incorporated into the ALIGNprogram (version 2.0), using a PAM120 weight residue table, a gap lengthpenalty of 12 and a gap penalty of 4. In addition, the percent identitybetween two amino acid sequences can be determined using the Needlemanand Wunsch (J. Mol, Biol. 48:444-453, 1970) algorithm which has beenincorporated into the GAP program in the GCG software package (availableat www.gcg.com), using either a Blossom 62 matrix or a PAM250 matrix,and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1,2, 3, 4, 5, or 6.

Other than percentage of sequence identity noted above, anotherindication that two nucleic acid sequences or polypeptides aresubstantially identical is that the polypeptide encoded by the firstnucleic acid is immunologically cross reactive with the antibodiesraised against the polypeptide encoded by the second nucleic acid, asdescribed below. Thus, a polypeptide is typically substantiallyidentical to a second polypeptide, for example, where the two peptidesdiffer only by conservative substitutions. Another indication that twonucleic acid sequences are substantially identical is that the twomolecules or their complements hybridize to each other under stringentconditions, as described below. Yet another indication that two nucleicacid sequences are substantially identical is that the same primers canbe used to amplify the sequence.

The term “isolated antibody” refers to an antibody that is substantiallyfree of other antibodies having different antigenic specificities (e.g.,an isolated antibody that specifically binds C5 is substantially free ofantibodies that specifically bind antigens other than C5). An isolatedantibody that specifically binds C5 may, however, have cross-reactivityto other antigens. Moreover, an isolated antibody may be substantiallyfree of other cellular material and/or chemicals.

The term “isotype” refers to the antibody class (e.g., IgM, IgE, IgGsuch as IgG1 or IgG4) that is provided by the heavy chain constantregion genes. Isotype also includes modified versions of one of theseclasses, where modifications have been made to after the Fc function,for example, to enhance or reduce effector functions or binding to Fcreceptors.

The term “Kassoc” or “Ka”, as used herein, is intended to refer to theassociation rate of a particular antibody-antigen interaction, whereasthe term “Kdis” or “Kd,” as used herein, is intended to refer to thedissociation rate of a particular antibody-antigen interaction. The term“K_(D)”, as used herein, is intended to refer to the dissociationconstant, which is obtained from the ratio of Kd to Ka (i.e. Kd/Ka) andis expressed as a molar concentration (M). K_(D) values for antibodiescan be determined using methods well established in the art. A methodfor determining the K_(D) of an antibody is by using surface plasmonresonance, or using a biosensor system such as a Biacore® system.

The terms “monoclonal antibody” or “monoclonal antibody composition” asused herein refer to a preparation of antibody molecules of singlemolecular composition. A monoclonal antibody composition displays asingle binding specificity and affinity for a particular epitope.

The term “nucleic acid” is used herein interchangeably with the term“polynucleotide” and refers to deoxyribonucleotides or ribonucleotidesand polymers thereof in either single- or double-stranded form. The termencompasses nucleic acids containing known nucleotide analogs ormodified backbone residues or linkages, which are synthetic, naturallyoccurring, and non-naturally occurring, which have similar bindingproperties as the reference nucleic acid, and which are metabolized in amanner similar to the reference nucleotides. Examples of such analogsinclude, without limitation, phosphorothioates, phosphoramidates, methylphosphonates, chiral-methyl phosphonates, 2-O-methyl ribonucleotides,peptide-nucleic acids (PNAs).

Unless otherwise indicated, a particular nucleic acid sequence alsoimplicitly encompasses conservatively modified variants thereof (e.g.,degenerate codon substitutions) and complementary sequences, as well asthe sequence explicitly indicated. Specifically, as detailed below,degenerate codon substitutions may be achieved by generating sequencesin which the third position of one or more selected (or all) codons issubstituted with mixed-base and/or deoxyinosine residues (Batzer et al.,Nucleic Acid Res. 19:5081, 1991; Ohtsuka et al., J. Biol. Chem.260:2605-2608, 1985; and Rossolini et al., Mol. Cell. Probes 8:91-98,1994).

The term “operably linked” refers to a functional relationship betweentwo or more polynucleotide (e.g., DNA) segments. Typically, it refers tothe functional relationship of a transcriptional regulatory sequence toa transcribed sequence. For example, a promoter or enhancer sequence isoperably linked to a coding sequence if it stimulates or modulates thetranscription of the coding sequence in an appropriate host cell orother expression system. Generally, promoter transcriptional regulatorysequences that are operably linked to a transcribed sequence arephysically contiguous to the transcribed sequence, i.e., they arecis-acting. However, some transcriptional regulatory sequences, such asenhancers, need not be physically contiguous or located in closeproximity to the coding sequences whose transcription they enhance.

As used herein, the term, “optimized” means that a nucleotide sequencehas been altered to encode an amino acid sequence using codons that arepreferred in the production cell or organism, generally a eukaryoticcell, for example, a cell of Pichia, a Chinese Hamster Ovary cell (CHO)or a human cell. The optimized nucleotide sequence is engineered toretain completely or as much as possible the amino acid sequenceoriginally encoded by the starting nucleotide sequence, which is alsoknown as the “parental” sequence. The optimized sequences herein havebeen engineered to have codons that are preferred in mammalian cells.However, optimized expression of these sequences in other eukaryoticcells or prokaryotic cells is also envisioned herein. The amino acidsequences encoded by optimized nucleotide sequences are also referred toas optimized.

The terms “polypeptide” and “protein” are used interchangeably herein torefer to a polymer of amino acid residues. The terms apply to amino acidpolymers in which one or more amino acid residue is an artificialchemical mimetic of a corresponding naturally occurring amino acid, aswell as to naturally occurring amino acid polymers and non-naturallyoccurring amino acid polymer. Unless otherwise indicated, a particularpolypeptide sequence also implicitly encompasses conservatively modifiedvariants thereof.

The term “recombinant human antibody”, as used herein, includes allhuman antibodies that are prepared, expressed, created or isolated byrecombinant means, such as antibodies isolated from an animal (e.g., amouse) that is transgenic or transchromosomal for human immunoglobulingenes or a hybridoma prepared therefrom, antibodies isolated from a hostcell transformed to express the human antibody, e.g., from atransfectoma, antibodies isolated from a recombinant, combinatorialhuman antibody library, and antibodies prepared, expressed, created orisolated by any other means that involve splicing of all or a portion ofa human immunoglobulin gene, sequences to other DNA sequences. Suchrecombinant human antibodies have variable regions in which theframework and CDR regions are derived from human germline immunoglobulinsequences. In certain embodiments, however, such recombinant humanantibodies can be subjected to in vitro mutagenesis (or, when an animaltransgenic for human Ig sequences is used, in vivo somatic mutagenesis)and thus the amino acid sequences of the VH and VL regions of therecombinant antibodies are sequences that, while derived from andrelated to human germline VH and VL sequences, may not naturally existwithin the human antibody germline repertoire in vivo.

The term “recombinant host cell” (or simply “host cell”) refers to acell into which a recombinant expression vector has been introduced. Itshould be understood that such terms are intended to refer not only tothe particular subject cell but to the progeny of such a cell. Becausecertain modifications may occur in succeeding generations due to eithermutation or environmental influences, such progeny may not, in fact, beidentical to the parent cell, but are still included within the scope ofthe term “host cell” as used herein.

The term “subject” includes human and non-human animals. Non-humananimals include all vertebrates, e.g., mammals and non-mammals, such asnon-human primates, sheep, dog, cow, chickens, amphibians, and reptiles.Except when noted, the terms “patient” or “subject” are used hereininterchangeably.

The term “treating” includes the administration of compositions orantibodies to prevent or delay the onset of the symptoms, complications,or biochemical indicia of a disease (e.g., AMD), alleviating thesymptoms or arresting or inhibiting further development of the disease,condition, or disorder. Treatment may be prophylactic (to prevent ordelay the onset of the disease, or to prevent the manifestation ofclinical or subclinical symptoms thereof) or therapeutic suppression oralleviation of symptoms after the manifestation of the disease.

The term “vector” is intended to refer to a polynucleotide moleculecapable of transporting another polynucleotide to which it has beenlinked. One type of vector is a “plasmid”, which refers to a circulardouble stranded DNA loop into which additional DNA segments may beligated. Another type of vector is a viral vector, wherein additionalDNA segments may be ligated into the viral genome. Certain vectors arecapable of autonomous replication in a host cell into which they areintroduced (e.g., bacterial vectors having a bacterial origin ofreplication and episomal mammalian vectors). Other vectors (e.g.,non-episomal mammalian vectors) can be integrated into the genome of ahost cell upon introduction into the host cell, and thereby arereplicated along with the host genome. Moreover, certain vectors arecapable of directing the expression of genes to which they areoperatively linked. Such vectors are referred to herein as “recombinantexpression vectors” (or simply, “expression vectors”). In general,expression vectors of utility in recombinant DNA techniques are often inthe form of plasmids. In the present specification, “plasmid” and“vector” may be used interchangeably as the plasmid is the most commonlyused form of vector. However, the invention is intended to include suchother forms of expression vectors, such as viral vectors (e.g.,replication defective retroviruses, adenoviruses and adeno-associatedviruses), which serve equivalent functions.

4. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows variable-region alignments of selected antibodies withtheir most closely related human germline sequences.

FIG. 2 shows a hemolytic assay in which human C5 is titrated into humanC5-depleted serum to determine C5 activity.

FIG. 3 shows titration of cynomolgus serum into human C5-depleted serumto determine optimal cynomolgus C5 concentration for alternative pathwayhemolytic assay.

FIG. 4 shows examples of classical pathway hemolytic assays with 20%human serum.

FIG. 5 shows example of alternative pathway hemolytic assays with 100 pMpurified human C5 added to human C5-depleted serum.

FIG. 6 shows examples of alternative pathway hemolytic assays with0.025% cynomolgus serum added to human C5-depleted serum.

FIG. 7 shows examples of classical pathway hemolytic assays (20% humanserum) with matured Fabs in comparison to their respective parentals.

FIG. 8 shows examples of classical pathway hemolytic assays (5%cynomolgus serum) with matured Fabs.

FIG. 9 shows affinity matured Fab characterization in alternativepathway hemolytic assay using 100 pM human C5 added to 20% humanC5-depleted serum.

FIG. 10 shows affinity matured Fab characterization in alternativepathway hemolytic assay using 20% human serum.

FIG. 11 shows affinity matured Fab characterization in alternativepathway hemolytic assay using 100 pM cynomolgus C5 added to 20% humanC5-depleted serum.

FIG. 12 shows characterization of germlined IgGs in classical pathwayhemolytic assay using 20% human serum.

FIG. 13 shows characterization of germlined IgGs in classical pathwayhemolytic assay using 5% cynomolgus serum.

FIG. 14 shows characterization of germlined IgGs in alternative pathwayhemolytic assay, 100 pM human C5.

FIG. 15 shows characterization of final germlined IgGs in alternativepathway hemolytic assay and C5a generation ELISA using 20% human serum.

FIG. 16 shows affinity matured Fab characterization in the C5a ELISAusing supernatant from 20% human serum hemolytic assays.

FIG. 17 shows specificity solution ELISA on human C3, C4, C5 andcynomolgus C5 testing antibody 7091 and its derivatives.

FIG. 18 shows serum stability assays (binding to human C5 in thepresence of 50% serum) with the Fabs.

FIG. 19 shows epitope binning of some affinity improved Fabs.

FIG. 20 shows an ELISA for antibody binding to mouse-human chimeric C5or human C5 to determine alpha chain versus beta chain binders. C5 waspresented by 5G1.1 to determine competition with 5G1.1.

FIG. 21 shows ELISA for testing alpha chain versus beta chain binderswith 5G1.1 capture.

FIG. 22 shows results of hemolytic assay for testing alpha chain versusbeta chain binders.

FIG. 23 show thermolysin proteolysis of parental Fabs at 37° C. (0, 30,60 and 90 minutes).

FIG. 24 show thermolysin proteolysis of parental Fabs at 55° C. (0, 30,60 and 90 minutes).

FIG. 25 shows thermolysin sensitivity of matured Fabs at 37° C.

FIG. 26 shows thermolysin sensitivity of matured Fabs at 55° C.

FIG. 27 shows examples of Fab inhibition of alternative pathway in MACdeposition assay.

5. DETAILED DESCRIPTION OF THE INVENTION

The present invention provides antibodies that specifically bind tocomplement C5 protein (e.g., human C5, cynomologus C5), pharmaceuticalcompositions, production methods, and methods of use of such antibodiesand compositions.

5.1. C5 ANTIBODIES

The present invention provides antibodies that specifically bind to C5(e.g., human C5, cynomologus C5). In some embodiments, the presentinvention provides antibodies that specifically bind to both human andcynomologus C5. Antibodies of the invention include, but are not limitedto, the human monoclonal antibodies, isolated as described, in theExamples (see Section 6 below).

The present invention provides antibodies that specifically bind a C5protein (e.g., human and/or cynomologus C5), said antibodies comprisinga VH domain having an amino acid sequence of SEQ ID NO: 7, 23, 39, 51,67, 79, 96, 108, 114, 121, 137, 151, 165, 179, 187, 201, 210, 218, 227,241, 253, 257, 273, 277, or 281. The present invention also providesantibodies that specifically bind to a C5 protein (e.g., human and/orcynomologus C5), said antibodies comprising a VH CDR having an aminoacid sequence of any one of the VH CDRs listed in Table 1, infra. Inparticular, the invention provides antibodies that specifically bind toa C5 protein (e.g., human and/or cynomologus C5), said antibodiescomprising (or alternatively, consisting of) one, two, three, four, fiveor more VH CDRs having an amino acid sequence of any of the VH CDRslisted in Table 1, infra.

The present invention provides antibodies that specifically bind to a C5protein (e.g., human and/or cynomologus C5), said antibodies comprisinga VL domain having an amino acid sequence of SEQ ID NO: 8, 24, 40, 52,68, 80, 90, 102, 122, 138, 152, 166, 180, 188, 202, 211, 219, 228, 242,261, 265, 269, 285, or 289. The present invention also providesantibodies that specifically bind to a C5 protein (e.g., human and/orcynomologus C5), said antibodies comprising a VL CDR having an aminoacid sequence of any one of the VL CDRs listed in Table 1, infra. Inparticular, the invention provides antibodies that specifically bind toa C5 protein (e.g., human and/or cynomologus C5), said antibodiescomprising (or alternatively, consisting of) one, two, three or more VLCDRs having an amino acid sequence of any of the VL CDRs listed in Table1, infra.

Other antibodies of the invention include amino acids that have beenmutated, yet have at least 60, 70, 80, 90 or 95 percent identity in theCDR regions with the CDR regions depicted in the sequences described inTable 1. In some embodiments, it includes mutant amino acid sequenceswherein no more than 1, 2, 3, 4 or 5 amino acids have been mutated inthe CDR regions when compared with the CDR regions depicted in thesequence described in Table 1.

The present invention also provides nucleic acid sequences that encodeVH, VL, the full length heavy chain, and the full length light chain ofthe antibodies that specifically bind to a C5 protein (e.g., humanand/or cynomologus C5). Such nucleic acid sequences can be optimized forexpression in mammalian cells (for example, Table 1 shows the optimizednucleic acid sequences for the heavy chain and light chain of antibodies8109, 8110, 8111, 8113, 8114, 8112, 8125, 8126, 8127, 8128, 8129, 8130,8131, 8132, and 8091).

TABLE 1 Examples of C5 Antibodies of the Present Invention and C5Proteins Sequence Identifier (SEQ ID NO:) or comments/details Antibody8109 CDRH1 1:SYAIS CDRH2 2:GIGPFFGTANYAQKFQG CDRH3 3:DTPYFDY CDRL14:SGDSIPNYYVY CDRL2 5:DDSNRPS CDRL3 6:QSFDSSLNAEV VH7:EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIGPFFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARDTPYFDYW GQGTLVTVSS VL8:SYELTQPLSVSVALGQTARITCSGDSIPNYYVYWYQQKPGQAPVLVIYDDSNRPSGIPERFSGSNSGNTATLTISRAQAGDEADYYCQSFDSSLNAEVFGGGTKLT VL Heavy chain9:EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIGPFFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARDTPYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVOGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPGK Light chain10:SYELTQPLSVSVALGQTARITCSGDSIPNYYVYWYQQKPGQAPVLVIYDDSNRPSGIPERFSGSNSGNTATLTISRAQAGDEADYYCQSFDSSLNAEVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTEC S PN encoding11:GAGGTGCAATTGGTTCAGTCTGGCGCGGAAGTG SEQ ID NO: 7AAAAAACCGGGCAGCAGCGTGAAAGTGAGCTGCAAAGCCTCCGGAGGCACTTTTTCTTCTTATGCCATTTCTTGGGTGCGCCAAGCCCCTGGGCAGGGTCTCGAGTGGATGGGCGGTATCGGTCCGTTTTTTGGCACTGCGAATTACGCGCAGAAGTTTCAGGGCCGGGTGACCATTACCGCGGATGAAAGCACCAGCACCGCGTATATGGAACTGAGCAGCCTGCGTAGCGAAGATACGGCCGTGTATTATTGCGCGCGTGATACTCCTTATTTTGATTATTGGGGC CAAGGCACCCTGGTGACGGTTAGCTCA PNencoding 12:TCCTATGAACTCACACAGCCCCTGAGCGTGAGC SEQ ID NO: 8GTGGCCCTGGGCCAGACCGCCCGGATCACCTGCTCCGGCGACAGCATCCCCAACTACTACGTGTACTGGTACCAGCAGAAGCCCGGCCAGGCCCCCGTGCTGGTGATCTACGACGACAGCAACCGGCCCAGCGGCATCCCCGAGCGGTTCAGCGGCAGCAACAGCGGCAACACCGCCACCCTGACCATTTCCAGAGCACAGGCAGGCGACGAGGCCGACTACTACTGCCAGAGCTTCGACAGCAGCCTGAACGCCGAGGTGTTCGGCGGAGGGACCAAGTTAACCGTC CTA PN encoding13:GAGGTGCAATTGGTTCAGTCTGGCGCGGAAGTG SEQ ID NO: 9AAAAAACCGGGCAGCAGCGTGAAAGTGAGCTGCAAAGCCTCCGGAGGCACTTTTTCTTCTTATGCCATTTCTTGGGTGCGCCAAGCCCCTGGGCAGGGTCTCGAGTGGATGGGCGGTATCGGTCCGTTTTTTGGCACTGCGAATTACGCGCAGAAGTTTCAGGGCCGGGTGACCATTACCGCGGATGAAAGCACCAGCACCGCGTATATGGAACTGAGCAGCCTGCGTAGCGAAGATACGGCCGTGTATTATTGCGCGCGTGATACTCCTTATTTTGATTATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCCTCCACCAAGGGTCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCAGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCT CCGGGTAAA PN encoding14:TCCTATGAACTCACACAGCCCCTGAGCGTGAGC SEQ ID NO: 10GTGGCCCTGGGCCAGACCGCCCGGATCACCTGCTCCGGCGACAGCATCCCCAACTACTACGTGTACTGGTACCAGCAGAAGCCCGGCCAGGCCCCCGTGCTGGTGATCTACGACGACAGCAACCGGCCCAGCGGCATCCCCGAGCGGTTCAGCGGCAGCAACAGCGGCAACACCGCCACCCTGACCATTTCCAGAGCACAGGCAGGCGACGAGGCCGACTACTACTGCCAGAGCTTCGACAGCAGCCTGAACGCCGAGGTGTTCGGCGGAGGGACCAAGTTAACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTACTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACC GTGGAGAAGACAGTGGCCCCTACAGAATGTTCAOptimized PN 15:GAGGTGCAGCTGGTGCAGAGCGGAGCCGAGGTG encodingAAGAAGCCCGGTAGCAGCGTCAAGGTGTCCTGCAAG SEQ ID NO: 9GCCAGCGGCGGCACCTTCAGCAGCTACGCCATCAGCTGGGTGCGGCAGGCCCCAGGCCAGGGCCTGGAGTGGATGGGCGGCATCGGCCCATTCTTCGGCACCGCCAACTACGCCCAGAAGTTCCAGGGCAGGGTCACCATCACCGCCGACGAGAGCACCAGCACCGCCTACATGGAGCTGTCCAGCCTGAGAAGCGAGGACACCGCCGTGTACTACTGCGCCAGAGACACCCCCTACTTCGACTACTGGGGCCAGGGCACCCTGGTGACCGTGAGCAGCGCTAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCTCCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCCCCCGAAGCTGCAGGCGGCCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGACGTGAGCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAGGTCTCCAACAAGGCCCTGCCTGCCCCCATCGAAAAGACCATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCTTCTCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTGTCA CCCGGCAAG Optimized PN16:AGCTACGAGCTGACCCAGCCCCTGAGCGTGAGC encodingGTGGCCCTGGGCCAGACCGCCAGGATCACCTGCAGC SEQ ID NO: 10GGCGACAGCATCCCCAACTACTACGTGTACTGGTATCAGCAGAAGCCCGGCCAGGCCCCCGTGCTGGTGATCTACGACGACAGCAACAGGCCCAGCGGCATCCCCGAGAGGTTCAGCGGCAGCAACAGCGGCAACACCGCCACCCTGACCATCAGCAGAGCCCAGGCCGGCGACGAGGCCGACTACTACTGCCAGAGCTTCGACAGCTCACTGAACGCCGAGGTGTTCGGCGGAGGGACCAAGCTGACCGTGCTGGGCCAGCCTAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACCCTGGTGTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACC GTGGAAAAGACCGTGGCCCCAACCGAGTGCAGCAntibody 8110 CDRH1 17:NYIS CDRH2 18:IIDPDDSYTEYSPSFQG CDRH3 19:YEYGGFDICDRL1 20:SGDNIGNSYVH CDRL2 21:KDNDRPS CDRL3 22:GTYDIESYV VH23:EVQLVQSGAEVKKPGESLKISCKGSGYSFTNYISWVRQMPGKGLEWMGIIDPDDSYTEYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYEYGGFDI WGQGTLVTVSS VL24:SYELTQPPSVSVAPGQTARISCSGDNIGNSYVHWYQQKPGQAPVLVIYKDNDRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCGTYDIESYVFGGGTKLTV L Heavy chain25:EVQLVQSGAEVKKPGESLKISCKGSGYSFTNYISWVRQMPGKGLEWMGIIDPDDSYTEYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYEYGGFDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPGK Light chain26:SYELTQPPSVSVAPGQTARISCSGDNIGNSYVHWYQQKPGQAPVLVIYKDNDRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCGTYDIESYVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSY LSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSPN encoding 27:GAGGTGCAATTGGTTCAGAGCGGCGCGGAAGTG SEQ ID NO: 23AAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTAATTATATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCACATTGATCCTGATGATTCTTATACTGAGTATTCTCCTTCTTTTCAGGGTCAGGTCACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATGAGTATGGTGGTTTTGATATTTGGGGCC AAGGCACCCTGGTGACGGTTAGCTCA PNencoding 28:AGTTACGAACTGACCCAGCCGCCTTCAGTGAGC SEQ ID NO: 24GTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATAATATTGGTAATTCTTATGTTCATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATGATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCGGTACTTATGATATVGAGTCTTAT GTGTTTGGCGGCGGCACGAAGTTAACCGTCCTAPN encoding 29:GAGGTGCAATTGGTTCAGAGCGGCGCGGAAGTG SEQ ID NO: 25AAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTAATTATATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTGATGATTCTTATACTGAGTATTCTCCTTCTTTTCAGGGTCAGGTCACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATGAGTATGGTGGTTTTGATATTTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCCTCCACCAAGGGTCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCAGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCT CCGGGTAAA PN encoding30:AGTTACGAACTGACCCAGCCGCCTTCAGTGAGC SEQ ID NO: 26GTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATAATATTGGTAATTCTTATGTTCATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATGATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCGGTACTTATGATATTGAGTCTTATGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAG AAGACAGTGGCCCCTACAGAATGTTCAOptimized PN 31:GAGGTGCAGCTGGTGCAGAGCGGAGCCGAGGTG encodingAAAAAGCCCGGTGAGAGCCTGAAGATCAGCTGCAAG SEQ ID NO: 25GGCAGCGGCTACAGCTTCACCAACTACATCAGCTGGGTGCGGCAGATGCCCGGCAAGGGCCTGGAGTGGATGGGCATCATCGACCCCGACGACAGCTACACCGAGTACAGCCCCAGCTTCCAGGGCCAGGTGACCATCAGCGCCGACAAGAGCATCAGCACCGCCTACCTGCAGTGGAGCAGCCTGAAGGCCAGCGACACCGCCATGTACTACTGCGCCAGATACGAGTACGGCGGCTTCGACATCTGGGGCCAGGGCACCCTGGTGACCGTCAGCTCAGCTAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCTCCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCCCCCGAAGCTGCAGGCGGCCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGACGTGAGCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAGGTCTCCAACAAGGCCCTGCCTGCCCCCATCGAAAAGACCATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCTTCTCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTGTCA CCCGGCAAG Optimized PN32:AGCTACGAGCTGACCCAGCCCCCCAGCGTGAGC encodingGTGGCCCCAGGCCAGACCGCCAGGATCAGCTGCAGC SEQ ID NO: 26GGCGACAACATCGGCAACAGCTACGTGCACTGGTATCAGCAGAAGCCCGGCCAGGCCCCCGTGCTGGTGATCTACAAGGACAACGACAGGCCCAGCGGCATCCCCGAGAGGTTCAGCGGCAGCAACTCCGGCAACACCGCCACCCTGACCATCAGCGGCACCCAGGCCGAGGACGAGGCCGACTACTACTGCGGCACCTACGACATCGAGTCATACGTGTTCGGCGGAGGGACCAAGCTGACCGTGCTGGGCCAGCCTAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACCCTGGTGTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGTGGAA AAGACCGTGGCCCCAACCGAGTGCAGCAntibody 8111 CDRH1 33:TSGGGVS CDRH2 34:NIDDADIKDYSPSLKS CDRH335:GPYGFDS CDRL1 36:TGTSSDIGTYNYVS CDRL2 37:DDSNRPS CDRL3 38:QSYDSQSIVVH 39:EVTLKESGPALVKPTQTLTLTCTFSGFSLSTSGGGVSWIRQPPGKALEWLANIDDADIKDYSPSLKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARGPYGFD SWGQGTLVTVSS VL40:ESALTQPASVSGSPGQSITISCTGTSSDIGTYNYVSWYQQHPGKAPKLMIYDDSNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCQSYDSQSIVFGGGTK LTVL Heavy chain41:EVTLKESGPALVKPTQTLTLTCTFSGFSLSTSGGGVSWIRQPPGKALEWLANIDDADIKOYSPSLKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARGPYGFDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKQYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPGK Light chain42:ESALTQPASVSGSPGQSITISCTGTSSDIGTYNYVSWYQQHPGKAPKLMIYDDSNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCQSYDSQSIVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTE CS PN encoding43:GAGGTGACATGAAAGAAAGCGGCCCGGCCCTGG SEQ ID NO: 39TGAAACCGACCCAAACCCTGACCCTGACCTGTACCTTTTCCGGATTTAGCCTGTCTACTTCTGGTGGTGGTGTGTCTTGGATTCGCCAGCCGCCTGGGAAAGCCCTCGAGTGGCTGGCTAATATTGATGATGCTGATATTAAGGATTATTCTCCTTCTCTTAAGTCTCGTCTGACCATTAGCAAAGATACTTCGAAAAATCAGGTGGTGCTGACTATGACCAACATGGACCCGGTGGATACGGCCACCTATTATTGCGCGCGTGGTCCTTATGGTTTTGATTCTTGGG GCCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding 44:GAAAGCGCACTGACCCAGCCAGCTCAGTGAGCG SEQ ID NO: 40GCTCACCAGGTCAGAGCATTACCATCTCGTGTACGGGTACTAGCAGCGATATTGGTACTTATAATTATGTGTCTTGGTACCAGCAGCATCCCGGGAAGGCGCCGAAACTTATGATTTATGATGATTCTAATCGTCCCTCAGGCGTGAGCAACCGTTTTAGCGGATCCAAAAGCGGCAACACCGCGAGCCTGACCATTAGCGGCCTGCAAGCGGAAGACGAAGCGGATTATTATTGCCAGTCTTATGATTCTCAGTCTATTGTGTTTGGCGGCGGCACGAAGTTAACCG TCCTA PN encoding45:GAGGTGACATTGAAAGAAAGCGGCCCGGCCCTG SEQ ID NO: 41GTGAAACCGACCCAAACCCTGACCCTGACCTGTACCTTTTCCGGATTTAGCCTGTCTACTTCTGGTGGTGGTGTGTCTTGGATTCGCCAGCCGCCTGGGAAAGCCCTCGAGTGGCTGGCTAATATTGATGATGCTGATATTAAGGATTATTCTCCTTCTCTTAAGTCTCGTCTGACCATTAGCAAAGATACTTCGAAAAATCAGGTGGTGCTGACTATGACCAACATGGACCCGGTGGATACGGCCACCTATTATTGCGCGCGTGGTCCTTATGGTTTTGATTCTTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCCTCCACCAAGGGTCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCAGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCGCGACAACATCGGCAACAGCTACGTGCACTGGTATCAGCAGAAGCCCGGCCAGGCCCCCGTGCTGGTGATCAGAAGAGCCTCTCCCTGT CTCCGGGTAAA PN encoding46:GAAAGCGCACTGACCCAGCCAGCTTCAGTGAGC SEQ ID NO: 42GGCTCACCAGGTCAGAGCATTACCATCTCGTGTACGGGTACTAGCAGCGATATTGGTACTTATAATTATGTGTCTTGGTACCAGCAGCATCCCGGGAAGGCGCCGAAACTTATGATTTATGATGATTCTAATCGTCCCTCAGGCGTGAGCAACCGTTTTAGCGGATCCAAAAGCGGCAACACCGCGAGCCTGACCATTAGCGGCCTGCAAGCGGAAGACGAAGCGGATTATTATTGCCAGTCTTATGATTCTCAGTCTATTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCA Optimized PN47:GAGGTGACCCTGAAGGAGAGCGGCCCAGCCCTG encodingGTGAAGCCCACCCAGACCCTGACCCTGACTTGCACC SEQ ID NO: 41TTCAGCGGCTTCAGCCTGAGCACCAGCGGAGGGGGCGTGAGCTGGATCAGGCAGCCCCCAGGTAAGGCCCTGGAGTGGCTGGCCAATATCGACGACGCCGATATCAAGGACTACAGCCCCAGCCTGAAGAGCAGGCTGACCATCAGCAAGGACACCAGCAAGAACCAGGTGGTGCTGACCATGACCAATATGGACCCCGTGGACACCGCCACCTACTACTGCGCCAGAGGCCCCTACGGCTTCGACAGCTGGGGCCAGGGCACCCTGGTGACCGTCAGCTCAGCTAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCTCCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCCCCCGAAGCTGCAGGCGGCCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGACGTGAGCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAGGTCTCCAACAAGGCCCTGCCTGCCCCCATCGAAAAGACCATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCTTCTCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTG TCACCCGGCAAG Optimized PN48:GAGAGCGCCCTGACCCAGCCCGCCAGCGTGAGC encodingGGCAGCCCAGGCCAGTCTATCACAATCAGCTGCACC SEQ ID NO: 42GGCACCTCCAGCGATATCGGCACCTACAACTACGTGAGCTGGTATCAGCAGCACCCCGGCAAGGCCCCCAAGCTGATGATCTACGACGACAGCAACAGGCCCAGCGGCGTGAGCAACAGGTTCAGCGGCAGCAAGAGCGGCAACACCGCCAGCCTGACAATCAGCGGCCTGCAGGCCGAGGACGAGGCCGACTACTACTGCCAGAGCTACGACAGCCAGTCAATCGTGTTCGGCGGAGGGACCAAGCTGACCGTGCTGGGCCAGCCTAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACCCTGGTGTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGTGGAAAAGACCGTGGCCCCAACCGAGTGCAGC Antibody 8113 CDRH1 SEQ ID NO: 17CDRH2 49:IIDPDDSYTRYSPSFQG CDRH3 SEQ ID NO: 19 CDRL1 SEQ ID NO: 20 CDRL2SEQ ID NO: 21 CDRL3 50:ATWGSEDQV VH 51:EVQLVQSGAEVKKPGESLKISCKGSGYSFTNYISWVRQMPGKGLEWMGIIDPDDSYTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYEYGGFDI WGQGTLVTVSS VL52:SYELTQPPSVSVAPGQTARISCSGDNIGNSYVHWYQQKPGQAPVLVIYKDNDRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCATWGSEDQVFGGGTKLTV L Heavy chain53:EVQLVQSGAEVKKPGESLKISCKGSGYSFTNYISWVRQMPGKGLEWMGIIDPDDSYTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYEYGGFDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPGK Light chain54:SYELTQPPSVSVAPGQTARISCSGDNIGNSYVHWYQQKPGQAPVLVIYKDNDRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCATWGSEDQVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSY LSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSPN encoding 55:GAGGTGCAATTGGTTCAGAGCGGCGCGGAAGTG SEQ ID NO: 51AAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTAATTATATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATCGATCCGGATGATAGCTATACCCGTTATTCTCCGAGCTTTCAGGGACAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATGAGTATGGTGGTTTTGATATTTGGGGC CAAGGCACCCTGGTGACGGTTAGCTCA PNencoding 56:AGTTACGAACTGACCCAGCCGCCTTCAGTGAGC SEQ ID NO: 52GTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATAATATTGGTAATTCTTATGTTCATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATGATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCGCTACTTGGGGTTCTGAGGATCAG GTGTTTGGCGGCGGCACGAAGTTAACCGTCCTAPN encoding 57:GAGGTGCAATTGGTTCAGAGCGGCGCGGAAGTG SEQ ID NO: 53AAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTAATTATATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATCGATCCGGATGATAGCTATACCCGTTATTCTCCGAGCTTTCAGGGACAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATGAGTATGGTGGTTTTGATATTTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCCTCCACCAAGGGTCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCAGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCT CCGGGTAAA PN encoding58:AGTTACGAACTGACCCAGCCGCCTTCAGTGAGC SEQ ID NO: 54GTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATAATATTGGTAATTCTTATGTTCATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATGATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCGCTACTTGGGGTTCTGAGGATCAGGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAG AAGACAGTGGCCCCTACAGAATGTTCAOptimized PN 59:GAGGTGCAGCTGGTGCAGAGCGGAGCCGAGGTG encodingAAAAAGCCCGGTGAGAGCCTGAAGATCAGCTGCAAG SEQ ID NO: 53GGCAGCGGCTACAGCTTCACCAACTACATCAGCTGGGTGCGGCAGATGCCCGGCAAGGGCCTGGAGTGGATGGGCATCATCGACCCCGACGACAGCTACACCAGGTACAGCCCCAGCTTCCAGGGCCAGGTGACCATCAGCGCCGACAAGAGCATCAGCACCGCCTACCTGCAGTGGAGCAGCCTGAAGGCCAGCGACACCGCCATGTACTACTGCGCCAGATACGAGTACGGCGGCTTCGACATCTGGGGCCAGGGCACCCTGGTGACCGTCAGCTCAGCTAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCTCCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCCCCCGAAGCTGCAGGCGGCCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGACGTGAGCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAGGTCTCCAACAAGGCCCTGCCTGCCCCCATCGAAAAGACCATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCTTCTCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTGTCA CCCGGCAAG Optimized PN60:AGCTACGAGCTGACCCAGCCCCCCAGCGTGAGC encodingGTGGCCCCAGGCCAGACCGCCAGGATCAGCTGCAGC SEQ ID NO: 54GGCGACAATATCGGCAACAGCTACGTGCACTGGTATCAGCAGAAGCCCGGCCAGGCCCCCGTGCTGGTGATCTACAAGGACAACGACAGGCCCAGCGGCATCCCCGAGAGGTTCAGCGGCAGCAACTCCGGCAACACCGCCACCCTGACAATCAGCGGCACCCAGGCCGAGGACGAGGCCGACTACTACTGCGCCACCTGGGGCTCAGAGGACCAGGTGTTCGGCGGAGGGACCAAGCTGACCGTGCTGGGCCAGCCTAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACCCTGGTGTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGTGGAA AAGACCGTGGCCCCAACCGAGTGCAGCAntibody 8114 CDRH1 61:SYYIG CDRH2 62:IIDPTDSQTAYSPSFQG CDRH363:YMMRGFDH CDRL1 64:SGDSLGDYYAY CDRL2 65:KDNNRPS CDRL3 66:QTWDTGESGV VH67:EVQLVQSGAEVKKPGESLKISCKGSGYSFTSYYIGWVRQMPGKGLEWMGIIDPTDSQTAYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYMMRGFD HWGQGTLVTVSS VL68:SYELTQPPSVSVAPGQTARISCSGDSLGDYYAYWYQQKPGQAPVLVIYKDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQTWOTGESGVFGGGTKLT VL Heavy chain69:EVQLVQSGAEVKKPGESLKISCKGSGYSFTSYYIGVWRQMPGKGLEWMGIIDPTDSQTAYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYMMRGFDHWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHE ALHNHYTQKSLSLSPGK Light chain70:SYELTQPPSVSVAPGQTARISCSGDSLGDYYAYWYQQKPGQAPVLVIYKDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQTWDTGESGVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS PN encoding71:GAGGTGCAATTGGTTCAGAGCGGCGCGGAAGTG SEQ ID NO: 67AAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTTCTTATTATATTGGTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTACTGATTCTCAGACTGCTTATTCTCCTTCTTTTCAGGGTCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATATGATGCGTGGTTTTGATCATTGG GGCCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding 72:AGTTACGAACTGACCCAGCCGCCTTCAGTGAGC SEQ ID NO: 68GTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTCTTGGTGATTATTATGCTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGACTTGGGATACTGGTGAGTCTGGTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTA PN encoding73:GAGGTGCAATTGGTTCAGAGCGGCGCGGAAGTG SEQ ID NO: 69AAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTTCTTATTATATTGGTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTACTGATTCTCAGACTGCTTATTCTCCTTCTTTTCAGGGTCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATATGATGCGTGGTTTTGATCATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCCTCCACCAAGGGTCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCAGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTG TCTCCGGGTAAA PN encoding74:AGTTACGAACTGACCCAGCCGCCTTCAGTGAGC SEQ ID NO: 70GTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTCTTGGTGATTATTATGCTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGACTTGGGATACTGGTGAGTCTGGTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTG GAGAAGACAGTGGCCCCTACAGAATGTTCAOptimized PN 75:GAGGTGCAGCTGGTGCAGAGCGGAGCCGAGGTG encodingAAAAAGCCCGGTGAGAGCCTGAAGATCAGCTGCAAG SEQ ID NO: 69GGCAGCGGCTACAGCTTCACCAGCTACTACATCGGCTGGGTGCGGCAGATGCCCGGCAAGGGCCTGGAGTGGATGGGCATCATCGACCCCACCGACAGCCAGACCGCCTACAGCCCCAGCTTCCAGGGCCAGGTGACCATCAGCGCCGACAAGAGCATCAGCACCGCCTACCTGCAGTGGAGCAGCCTGAAGGCCAGCGACACCGCCATGTACTACTGCGCCCGGTACATGATGAGGGGCTTCGACCACTGGGGTCAGGGCACCCTGGTGACCGTCAGCTCAGCTAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCTCCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCCCCCGAAGCTGCAGGCGGCCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGACGTGAGCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAGGTCTCCAACAAGGCCCTGCCTGCCCCCATCGAAAAGACCATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCTTCTCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTG TCACCCGGCAAG Optimized PN76:AGCTACGAGCTGACCCAGCCCCCCAGCGTGAGC encodingGTGGCCCCAGGCCAGACCGCCAGGATCAGCTGCAGC SEQ ID NO: 70GGCGACAGCCTGGGCGACTACTACGCCTACTGGTATCAGCAGAAGCCCGGCCAGGCCCCCGTGCTGGTGATCTACAAGGACAACAACAGGCCCAGCGGCATCCCCGAGAGGTTCAGCGGCAGCAACAGCGGCAACACCGCCACCCTGACAATCAGCGGCACCCAGGCCGAGGACGAGGCCGACTACTACTGCCAGACCTGGGACACCGGCGAGTCAGGCGTGTTCGGCGGAGGGACCAAGCTGACCGTGCTGGGTCAGCCTAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACCCTGGTGTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGTG GAAAAGACCGTGGCCCCAACCGAGTGCAGCAntibody 8112 CDRH1 SEQ ID NO: 61 CDRH2 77:IIDPSOSHTTYSPSFQG CDRH3 SEQID NO: 63 CDRL1 SEQ ID NO: 64 CDRL2 SEQ ID NO: 65 CDRL3 78:QTWDILPHGLVVH 79:EVQLVQSGAEVKKPGESLKISCKGSGYSFTSYYIGVWRQMPGKGLEWMGIIDPSDSHTTYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYMMRGFD HWGQGTLVTVSS VL80:SYELTQPPSVSVAPGQTARISCSGDSLGDYYAYWYQQKPGQAPVLVIYKDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQTWDILPHGLVFGGGTKL TVL Heavy chain81:EVQLVQSGAEVKKPGESLKISCKGSGYSFTSYYIGVWRQMPGKGLEWMGIIDPSDSHTTYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYMMRGFDHWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHE ALHNHYTQKSLSLSPGK Light chain82:SYELTQPPSVSVAPGQTARISCSGDSLGDYYAYWYQQKPGQAPVLVIYKDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQTWDILPHGLVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTEC S PN encoding83:GAGGTGCAATTGGTTCAGAGCGGCGCGGAAGTG SEQ ID NO: 79AAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTTCTTATTATATTGGTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATCGATCCGTCTGATAGCCATACCACTTATTCTCCGAGCTTTCAGGGCCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATATGATGCGTGGTTTTGATCATTGG GGCCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding 84:AGTTACGAACTGACCCAGCCGCCTTCAGTGAGC SEQ ID NO: 80GTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTCTTGGTGATTATTATGCTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGACTTGGGATATTCTTCCTCATGGTCTTGTGTTTGGCGGCGGCACGAAGTTAACCGTC CTA PN encoding85:GAGGTGCAATTGGTTCAGAGCGGCGCGGAAGTG SEQ ID NO: 81AAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTTCTTATTATATTGGTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATCGATCCGTCTGATAGCCATACCACTTATTCTCCGAGCTTTCAGGGCCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATATGATGCGTGGTTTTGATCATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCCTCCACCAAGGGTCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCAGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTG TCTCCGGGTAAA PN encoding86:AGTTACGAACTGACCCAGCCGCCTTCAGTGAGC SEQ ID NO: 82GTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTCTTGGTGATTATTATGCTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGACTTGGGATATTCTTCCTCATGGTCTTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACC GTGGAGAAGACAGTGGCCCCTACAGAATGTTCAOptimized PN 87:GAGGTGCAGCTGGTGCAGAGCGGAGCCGAGGTG encodingAAAAAGCCCGGTGAGAGCCTGAAGATCAGCTGCAAG SEQ ID NO: 81GGCAGCGGCTACAGCTTCACCAGCTACTACATCGGCTGGGTGCGGCAGATGCCCGGCAAGGGCCTGGAGTGGATGGGCATTATCGATCCGTCTGATAGCCATACCACTTATTCTCCGAGCTTTCAGGGCCAGGTGACCATCAGCGCCGACAAGAGCATCAGCACCGCCTACCTGCAGTGGAGCAGCCTGAAGGCCAGCGACACCGCCATGTACTACTGCGCCCGGTACATGATGAGGGGCTTCGACCACTGGGGTCAGGGCACCCTGGTGACCGTCAGCTCAGCTAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCTCCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCCCCCGAAGCTGCAGGCGGCCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGACGTGAGCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAGGTCTCCAACAAGGCCCTGCCTGCCCCCATCGAAAAGACCATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCTTCTCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTG TCACCCGGCAAG Optimized PN88:AGCTACGAGCTGACCCAGCCCCCCAGCGTGAGC encodingGTGGCCCCAGGCCAGACCGCCAGGATCAGCTGCAGC SEQ ID NO: 82GGCGACAGCCTGGGCGACTACTACGCCTACTGGTATCAGCAGAAGCCCGGCCAGGCCCCCGTGCTGGTGATCTACAAGGACAACAACAGGCCCAGCGGCATCCCCGAGAGGTTCAGCGGCAGCAACAGCGGCAACACCGCCACCCTGACAATCAGCGGCACCCAGGCCGAGGACGAGGCCGACTACTACTGCCAGACTTGGGATATTCTTCCTCATGGTCTTGTGTTCGGCGGAGGGACCAAGCTGACCGTGCTGGGTCAGCCTAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACCCTGGTGTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACC GTGGAAAAGACCGTGGCCCCAACCGAGTGCAGCAntibody 8125 CDRH1 SEQ ID NO: 61 CDRH2 SEQ ID NO: 77 CDRH3 SEQ ID NO:63 CDRL1 SEQ ID NO: 64 CDRL2 SEQ ID NO: 65 CDRL3 89:QAWTDSPTGLV VH SEQID NO: 79 VL 90:SYELTQPPSVSVAPGQTARISCSGDSLGDYYAYWYQQKPGQAPVLVIYKDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQAWTDSPTGLVFGGGTKL TVL Heavy chain SEQ ID NO: 81 Lightchain 91:SYELTQPPSVSVAPGQTARISCSGDSLGDYYAYWYQQKPGQAPVLVIYKDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQAWTDSPTGLVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTEC S PN encoding SEQ ID NO: 83 SEQ IDNO: 79 PN encoding 92:AGTTACGAACTGACCCAGCCGCCTTCAGTGAGC SEQ ID NO: 90GTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTCTTGGTGATTATTATGCTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGGCTTGGACTGATTCTCCTACTGGTCTTGTGTTTGGCGGCGGCACGAAGTTAACCGTC CTA PN encoding SEQ ID NO: 85 SEQID NO: 81 PN encoding 93:AGTTACGAACTGACCCAGCCGCCTTCAGTGAGC SEQ ID NO: 91GTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTCTTGGTGATTATTATGCTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGGCTTGGACTGATTCTCCTACTGGTCTTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACC GTGGAGAAGACAGTGGCCCCTACAGAATGTTCAOptimized PN SEQ ID NO:87 encoding SEQ ID NO: 81 Optimized PN94:AGCTACGAGCTGACCCAGCCCCCCAGCGTGAGC encodingGTGGCCCCAGGCCAGACCGCCAGGATCAGCTGCAGC SEQ ID NO: 91GGCGACAGCCTGGGCGACTACTACGCCTACTGGTATCAGCAGAAGCCCGGCCAGGCCCCCGTGCTGGTGATCTACAAGGACAACAACAGGCCCAGCGGCATCCCCGAGAGGTTCAGCGGCAGCAACAGCGGCAACACCGCCACCCTGACAATCAGCGGCACCCAGGCCGAGGACGAGGCCGACTACTACTGCCAGGCTTGGACTGATTCTCCTACTGGTCTTGTGTTCGGCGGAGGGACCAAGCTGACCGTGCTGGGTCAGCCTAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACCCTGGTGTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACC GTGGAAAAGACCGTGGCCCCAACCGAGTGCAGCAntibody 8125 CDRH1 SEQ ID NO: 61 CDRH2 SEQ ID NO: 62 CDRH3 SEQ ID NO:63 CDRL1 SEQ ID NO: 64 CDRL2 SEQ ID NO: 65 CDRL3 SEQ ID NO: 89 VH SEQ IDNO: 67 VL SEQ ID NO: 90 Heavy chain SEQ ID NO: 69 Light chain SEQ ID NO:91 PN encoding SEQ ID NO: 71 SEQ ID NO: 79 PN encoding SEQ ID NO: 92 SEQID NO: 90 PN encoding SEQ ID NO: 73 SEQ ID NO: 81 PN encoding SEQ ID NO:93 SEQ ID NO: 91 Optimized PN SEQ ID NO: 75 encoding SEQ ID NO: 81Optimized PN SEQ ID NO: 94 encoding SEQ ID NO: 91 Antibody 8127 CDRH1SEQ ID NO: 61 CDRH2 95:IIDPTDSYTVYSPSFQG CDRH3 SEQ ID NO: 63 CDRL1 SEQID NO: 64 CDRL2 SEQ ID NO: 65 CDRL3 SEQ ID NO: 89 VH96:EVQLVQSGAEVKKPGESLKISCKGSGYSFTSYYIGVVVRQMPGKGLEWMGIIDPTDSYTVYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYMMRGF DHWGQGTLVTVSS VL SEQ ID NO: 90Heavy chain 97:EVQLVQSGAEVKKPGESLKISCKGSGYSFTSYYIGVWRQMPGKGLEWMGIIDPTDSYTVYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYMMRGFDHWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHE ALHNHYTQKSLSLSPGK Light chain SEQID NO: 91 PN encoding 98:GAGGTGCAATTGGTTCAGAGCGGCGCGGAAGTG SEQ ID NO: 96AAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTTCTTATTATATTGGTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTACTGATTCTTATACTGTTTATTCTCCTTCTTTTCAGGGTCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATATGATGCGTGGTTTTGATCATTGG GGCCAAGGCACCCTGGTGACGGTTAGCTCAGC PNencoding SEQ ID NO: 92 SEQ ID NO: 90 PN encoding99:GAGGTGCAATTGGTTCAGAGCGGCGCGGAAGTG SEQ ID NO: 97AAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTTCTTATTATATTGGTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTACTGATTCTTATACTGTTTATTCTCCTTCTTTTCAGGGTCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATATGATGCGTGGTTTTGATCATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCGCCTCCACCAAGGGTCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCAGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCC TGTCTCCGGGTAAA PN encoding SEQ IDNO: 93 SEQ ID NO: 91 Optimized PN 100:GAGGTGCAGCTGGTGCAGAGCGGAGCCGAGGTencoding GAAAAAGCCCGGTGAGAGCCTGAAGATCAGCTGCAA SEQ ID NO: 97GGGCAGCGGCTACAGCTTCACCAGCTACTACATCGGCTGGGTGCGGCAGATGCCCGGCAAGGGCCTGGAGTGGATGGGCATTATTGATCCTACTGATTCTTATACTGTTTATTCTCCTTCTTTTCAGGGTCAGGTGACCATCAGCGCCGACAAGAGCATCAGCACCGCCTACCTGCAGTGGAGCAGCCTGAAGGCCAGCGACACCGCCATGTACTACTGCGCCCGGTACATGATGAGGGGCTTCGACCACTGGGGTCAGGGCACCCTGGTGACCGTCAGCTCAGCTAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCTCCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCCCCCGAAGCTGCAGGCGGCCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGACGTGAGCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAGGTCTCCAACAAGGCCCTGCCTGCCCCCATCGAAAAGACCATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCTTCTCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCT GTCACCCGGCAAG Optimized PN SEQ IDNO: 94 encoding SEQ ID NO: 91 Antibody 8128 CDRH1 SEQ ID NO: 17 CDRH2SEQ ID NO: 49 CDRH3 SEQ ID NO: 19 CDRL1 SEQ ID NO: 20 CDRL2 SEQ ID NO:21 CDRL3 101:STWDIEPTYV VH SEQ ID NO: 51 VL102:SYELTQPPSVSVAPGQTARISCSGDNIGNSYVHWYQQKPGQAPVLVIYKDNDRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCSTWDIEPTYVFGGGTKL TVL Heavy chain SEQ ID NO: 53 Lightchain 103:SYELTQPPSVSVAPGQTARISCSGDNIGNSYVHWYQQKPGQAPVLVIYKDNDRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCSTWDIEPTYVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTEC S PN encoding SEQ ID NO: 55 SEQ IDNO: 51 PN encoding 104:AGTTACGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 102CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATAATATTGGTAATTCTTATGTTCATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATGATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCTCTACTTGGGATATTGAGCCTACTTATGTGTTTGGCGGCGGCACGAAGTTAACCGTCCT A PN encoding SEQ ID NO: 57 SEQ IDNO: 53 PN encoding 105:AGTTACGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 103CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATAATATTGGTAATTCTTATGTTCATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATGATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCTCTACTTGGGATATTGAGCCTACTTATGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGT GGAGAAGACAGTGGCCCCTACAGAATGTTCAOptimized PN SEQ ID NO: 59 encoding SEQ ID NO: 53 Optimized PN106:AGCTACGAGCTGACCCAGCCCCCCAGCGTGAG encodingCGTGGCCCCAGGCCAGACCGCCAGGATCAGCTGCAG SEQ ID NO: 103CGGCGACAATATCGGCAACAGCTACGTGCACTGGTATCAGCAGAAGCCCGGCCAGGCCCCCGTGCTGGTGATCTACAAGGACAACGACAGGCCCAGCGGCATCCCCGAGAGGTTCAGCGGCAGCAACTCCGGCAACACCGCCACCCTGACAATCAGCGGCACCCAGGCCGAGGACGAGGCCGACTACTACTGCTCTACTTGGGATATTGAGCCTACTTATGTGTTCGGCGGAGGGACCAAGCTGACCGTGCTGGGCCAGCCTAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACCCTGGTGTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGT GGAAAAGACCGTGGCCCCAACCGAGTGCAGCAntibody 8129 CDRH1 SEQ ID NO: 17 CDRH2 107:IIDPQDSYTEYSPSFQG CDRH3 SEQID NO: 19 CDRL1 SEQ ID NO: 20 CDRL2 SEQ ID NO: 21 CDRL3 SEQ ID NO: 22 VH108:EVQLVQSGAEVKKPGESLKISCKGSGYSFTNYISWVRQMPGKGLEWMGIIDPQDSYTEYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYEYGGFD IWGQGTLVTVSS VL SEQ ID NO: 24 Heavychain 109:EVQLVQSGAEVKKPGESLKISCKGSGYSFTNYISWVRQMPGKGLEWMGIIDPQDSYTEYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYEYGGFDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHE ALHNHYTQKSLSLSPGK Light chain SEQID NO: 26 PN encoding 110:GAGGTGCAATTGGTVCAGAGCGGCGCGGAAGT SEQ ID NO:108 GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTAATTATATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTCAGGATTCTTATACTGAGTATTCTCCTTCTTTTCAGGGTCAGGTCACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATGAGTATGGTGGTTTTGATATTTGGGG CCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding SEQ ID NO: 28 SEQ ID NO: 24 PN encoding111:GAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO 109GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTAATTATATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTCAGGATTCTTATACTGAGTATTCTCCTTCTTTTCAGGGTCAGGTCACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATGAGTATGGTGGTTTTGATATTTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCCTCCACCAAGGGTCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCAGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTC TCCGGGTAAA PN encoding SEQ ID NO:30 SEQ ID NO: 26 Optimized PN 112:GAGGTGCAGCTGGTGCAGAGCGGAGCCGAGGTencoding GAAAAAGCCCGGTGAGAGCCTGAAGATCAGCTGCAA SEQ ID NO: 109GGGCAGCGGCTACAGCTTCACCAACTACATCAGCTGGGTGCGGCAGATGCCCGGCAAGGGCCTGGAGTGGATGGGCATCATCGACCCCCAGGACAGCTACACCGAGTACAGCCCCAGCTTCCAGGGCCAGGTGACCATCAGCGCCGACAAGAGCATCAGCACCGCCTACCTGCAGTGGAGCAGCCTGAAGGCCAGCGACACCGCCATGTACTACTGCGCCAGATACGAGTACGGCGGCTTCGACATCTGGGGCCAGGGCACCCTGGTGACCGTCAGCTCAGCTAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCTCCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCCCCCGAAGCTGCAGGCGGCCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGACGTGAGCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAGGTCTCCAACAAGGCCCTGCCTGCCCCCATCGAAAAGACCATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCTTCTCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTGTC ACCCGGCAAG Optimized PN SEQ ID NO:32 encoding SEQ ID NO: 26 Antibody 8130 CDRH1 SEQ ID NO: 17 CDRH2 SEQ IDNO: 107 CDRH3 SEQ ID NO: 19 CDRL1 SEQ ID NO: 20 CDRL2 SEQ ID NO: 21CDRL3 SEQ ID NO: 101 VH SEQ ID NO: 108 VL SEQ ID NO: 102 Heavy chain SEQID NO: 109 Light chain SEQ ID NO: 103 PN encoding SEQ ID NO: 110 SEQ IDNO: 108 PN encoding SEQ ID NO: 104 SEQ ID NO: 102 PN encoding SEQ ID NO:111 SEQ ID NO: 109 PN encoding SEQ ID NO: 105 SEQ ID NO: 103 OptimizedPN SEQ ID NO: 112 encoding SEQ ID NO: 109 Optimized PN SEQ ID NO: 106encoding SEQ ID NO: 103 Antibody 8131 CDRH1 SEQ ID NO: 17 CDRH2113:IIDPEDSHTEYSPSFQG CDRH3 SEQ ID NO: 19 CDRL1 SEQ ID NO: 20 CDRL2 SEQID NO: 21 CDRL3 SEQ ID NO: 22 VH 114:EVQLVQSGAEVKKPGESLKISCKGSGYSFTNYISWVRQMPGKGLEWMGIIDPEDSHTEYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYEYGGFD IWGQGTLVTVSS VL SEQ ID NO: 24 Heavychain 115:EVQLVQSGAEVKKPGESLKISCKGSGYSFTNYISWVRQMPGKGLEWMGIIDPEDSHTEYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYEYGGFDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPGK Light chain SEQ IDNO: 26 PN encoding 116:GAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 114GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTAATTATATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTGAGGATTCTCATACTGAGTATTCTCCTTCTTTTCAGGGTCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATGAGTATGGTGGTTTTGATATTTGGGG CCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding SEQ ID NO: 28 SEQ ID NO: 24 PN encoding117:GAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 115GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTAATTATATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTGAGGATTCTCATACTGAGTATTCTCCTTCTTTTCAGGGTCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATGAGTATGGTGGTTTTGATATTTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCCTCCACCAAGGGTCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCAGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTC TCCGGGTAAA PN encoding SEQ ID NO:30 SEQ ID NO: 26 Optimized PN 118:GAGGTGCAGCTGGTGCAGAGCGGAGCCGAGGTencoding GAAAAAGCCCGGTGAGAGCCTGAAGATCAGCTGCAA SEQ ID NO: 115GGGCAGCGGCTACAGCTTCACCAACTACATCAGCTGGGTGCGGCAGATGCCCGGCAAGGGCCTGGAGTGGATGGGCATCATCGACCCCGAGGACAGCCATACCGAGTACAGCCCCAGCTTCCAGGGCCAGGTGACCATCAGCGCCGACAAGAGCATCAGCACCGCCTACCTGCAGTGGAGCAGCCTGAAGGCCAGCGACACCGCCATGTACTACTGCGCCAGATACGAGTACGGCGGCTTCGACATCTGGGGCCAGGGCACCCTGGTGACCGTCAGCTCAGCTAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCTCCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCCCCCGAAGCTGCAGGCGGCCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGACGTGAGCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAGGTCTCCAACAAGGCCCTGCCTGCCCCCATCGAAAAGACCATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCTTCTCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTGTC ACCCGGCAAG Optimized PN SEQ ID NO:32 encoding SEQ ID NO: 26 Antibody 8132 CDRH1 SEQ ID NO: 17 CDRH2 SEQ IDNO: 113 CDRH3 SEQ ID NO: 19 CDRL1 SEQ ID NO: 20 CDRL2 SEQ ID NO: 21CDRL3 SEQ ID NO: 101 VH SEQ ID NO: 114 VL SEQ ID NO: 102 Heavy chain SEQID NO: 115 Light chain SEQ ID NO: 103 PN encoding SEQ ID NO: 116 SEQ IDNO: 114 PN encoding SEQ ID NO: 104 SEQ ID NO: 102 PN encoding SEQ ID NO:117 SEQ ID NO: 115 PN encoding SEQ ID NO: 105 SEQ ID NO: 103 OptimizedPN SEQ ID NO: 11S encoding SEQ ID NO: 115 Optimized PN SEQ ID NO: 106encoding SEQ ID NO: 103 Antibody 8091 CDRH1 SEQ ID NO: 1 CDRH2119:NIGPFFGIANYAQKFQG CDRH3 SEQ ID NO: 3 CDRL1 SEQ ID NO: 4 CDRL2 SEQ IDNO: 5 CDRL3 120:QTYDDGSTAEV VH 121:QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGNIGPFFGIANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARDTPYFD YWGQGTLVTVSS VL122:DIELTQPPSVSVAPGQTARISCSGDSIPNYYVYWYQQKPGQAPVLVIYDDSNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQTYDDGSTAEVFGGGTK LTVL Heavy chain123:QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISVWRQAPGQGLEWMGNIGPFFGIANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARDTPYFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRWSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHN HYTQKSLSLSPGK Light chain124:DIELTQPPSVSVAPGQTARISCSGDSIPNYYVYWYQQKPGQAPVLVIYDDSNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQTYDDGSTAEVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTE CS PN encoding125:CAGGTGCAATTGGTTCAGTCTGGCGCGGAAGT SEQ ID NO: 121GAAAAAACCGGGCAGCAGCGTGAAAGTGAGCTGCAAAGCCTCCGGAGGCACTTTTTCTTCTTATGCCATTTCTTGGGTGCGCCAAGCCCCTGGGCAGGGTCTCGAGTGGATGGGCAATATCGGTCCGTTTTTTGGCATTGCGAATTACGCGCAGAAGTTTCAGGGCCGGGTGACCATTACCGCGGATGAAAGCACCAGCACCGCGTATATGGAACTGAGCAGCCTGCGTAGCGAAGATACGGCCGTGTATTATTGCGCGCGTGATACTCCTTATTTTGATTATTGGGG CCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding 126:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 122CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTATTCCTAATTATTATGTTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATGATGATTCTAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGACTTATGATGATGGTTCTACTGCTGAGGTGTTTGGCGGCGGCACGAAGTTAACCGT TCTT PN encoding127:CAGGTGCAATTGGTTCAGTCTGGCGCGGAAGT SEQ ID NO: 123GAAAAAACCGGGCAGCAGCGTGAAAGTGAGCTGCAAAGCCTCCGGAGGCACTTTTTCTTCTTATGCCATTTCTTGGGTGCGCCAAGCCCCTGGGCAGGGTCTCGAGTGGATGGGCAATATCGGTCCGTTTTTTGGCATTGCGAATTACGCGCAGAAGTTTCAGGGCCGGGTGACCATTACCGCGGATGAAAGCACCAGCACCGCGTATATGGAACTGAGCAGCCTGCGTAGCGAAGATACGGCCGTGTATTATTGCGCGCGTGATACTCCTTATTTTGATTATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCTTCCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCTGCAGCAGAAGCACCAGCGAGAGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGAGCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTGGACCACAAGCCCAGCAACACCAAGGTGGACAAGACCGTGGAGCGGAAGTGCTGCGTGGAGTGCCCCCCCTGCCCTGCCCCTCCTGTGGCCGGACCCTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCCGGACCCCCGAGGTGACCTGCGTGGTGGTGGACGTGAGCCACGAGGACCCCGAGGTGCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCCGGGAGGAACAGTTCAACAGCACCTTCCGGGTGGTGTCCGTGCTGACCGTGGTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTGCCCCCATCGAGAAAACCATCAGCAAGACAAAGGGCCAGCCCAGGGAACCCCAGGTGTACACCCTGCCCCCCAGCCGGGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCCATGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACAGTGGACAAGAGCCGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTA CACCCAGAAGAGCCTGAGCCTGTCCCCCGGCAAAPN encoding 128:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 124CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTATTCCTAATTATTATGTTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATGATGATTCTAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGACTTATGATGATGGTTCTACTGCTGAGGTGTTTGGCGGCGGCACGAAGTTAACCGTTCTTGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCAC CGTGGAGAAGACAGTGGCCCCTACAGAATGTTCAOptimized PN 129:CAGGTGCAGCTGGTGCAGTCCGGCGCCGAGGT encodingGAAGAAGCCCGGCTCCTCCGTGAAGGTGTCCTGCAA SEQ ID NO: 123GGCCTCCGGCGGCACCTTCTCCTCCTACGCCATCTCCTGGGTGCGGCAGGCCCCCGGCCAGGGCCTGGAGTGGATGGGCAACATCGGCCCCTTCTTCGGCATCGCCAACTACGCCCAGAAGTTCCAGGGCCGGGTGACCATCACCGCCGACGAGTCCACCTCCACCGCCTACATGGAGCTGTCCTCCCTGCGGTCCGAGGACACCGCCGTGTACTACTGCGCCCGGGACACCCCCTACTTCGACTACTGGGGCCAGGGCACCCTGGTGACCGTGTCCTCCGCCTCCACCAAGGGCCCCTCCGTGTTCCCCCTGGCCCCCTGCTCCCGGTCCACCTCCGAGTCCACCGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACTCCGGCGCCCTGACCTCCGGCGTGCACACCTTCCCCGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTGACCGTGCCCTCCTCCAACTTCGGCACCCAGACCTACACCTGCAACGTGGACCACAAGCCCTCCAACACCAAGGTGGACAAGACCGTGGAGCGGAAGTGCTGCGTGGAGTGCCCCCCCTGCCCCGCCCCCCCCGTGGCCGGCCCCTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCCGAGGTGACCTGCGTGGTGGTGGACGTGTCCCACGAGGACCCCGAGGTGCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCCGGGAGGAGCAGTTCAACTCCACCTTCCGGGTGGTGTCCGTGCTGACCGTGGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCCGCCCCCATCGAGAAGACCATCTCCAAGACCAAGGGCCAGCCCCGGGAGCCCCAGGTGTACACCCTGCCCCCCTCCCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCTCCGACATCGCCGTGGAGTGGGAGTCCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCCATGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCAAGCTGACCGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTA CACCCAGAAGTCCCTGTCCCTGTCCCCCGGCAAGOptimized PN 130:GACATCGAGCTGACCCAGCCCCCCTCCGTGTC encodingCGTGGCCCCCGGCCAGACCGCCCGGATCTCCTGCTC SEQ ID NO: 124CGGCGACTCCATCCCCAACTACTACGTGTACTGGTACCAGCAGAAGCCCGGCCAGGCCCCCGTGCTGGTGATCTACGACGACTCCAACCGGCCCTCCGGCATCCCCGAGCGGTTCTCCGGCTCCAACTCCGGCAACACCGCCACCCTGACCATCTCCGGCACCCAGGCCGAGGACGAGGCCGACTACTACTGCCAGACCTACGACGACGGCTCCACCGCCGAGGTGTTCGGCGGCGGCACCAAGCTGACCGTGCTGGGCCAGCCTAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACCCTGGTGTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCAC CGTGGAAAAGACCGTGGCCCCAACCGAGTGCAGCAntibody 6525 CDRH1 131:SYWIS CDRH2 132:IIDPDDSKTNYSPSFQG CDRH3133:RSYYPMDY CDRL1 134:TGTSSDVVGVYNFVS CDRL2 135:YVDNRPS CDRL3136:QSFDGFGIDMV VH 137:QVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWISWVRQMPGKGLEWMGIIDPDDSKTNYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARRSYYPM DYWGQGTLVTVSS VL138:DIALTQPASVSGSPGQSITISCTGTSSDWGVYNFVSWYQQHPGKAPKLMIYYVDNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCQSFDGFGIDMVFGG GTKLTVL Heavy chain139:QVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWISWVRQMPGKGLEWMGIIDPDDSKTNYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARRSYYPMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KKVEPKSX (X can be G, EF or CEF)Light chain 140:DIALTQPASVSGSPGQSITISCTGTSSDWGVYNFVSWYQQHPGKAPKLMIYYVDNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCQSFDGFGIDMVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVA PTEX (X can be CS or A) PN encoding141:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 137GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTTCTTATTGGATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATCGATCCGGATGATAGCAAGACCAATTATTCTCCGAGCTTTCAGGGCCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTCGTTCTTATTATCCTATGGATTATTG GGGCCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding 142:GATATCGCACTGACCCAGCCAGCTTCAGTGAG SEQ ID NO: 138CGGCTCACCAGGTCAGAGCATTACCATCTCGTGTACGGGTACTAGCAGCGATGTTGTTGGTGTTTATAATTTTGTGTCTTGGTACCAGCAGCATCCCGGGAAGGCGCCGAAACTTATGATTTATTATGTTGATAATCGTCCCTCAGGCGTGAGCAACCGTTTTAGCGGATCCAAAAGCGGCAACACCGCGAGCCTGACCATTAGCGGCCTGCAAGCGGAAGACGAAGCGGATTATTATTGCCAGTCTTTTGATGGTTTTGGTATTGATATGGTGTTTGGCGGCGGCAC GAAGTTAACCGTTCTT PN encoding143:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 139GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTTCTTATTGGATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATCGATCCGGATGATAGCAAGACCAATTATTCTCCGAGCTTTCAGGGCCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTCGTTCTTATTATCCTATGGATTATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCGTCGACCAAAGGTCCAAGCGTGTTTCCGCTGGCTCCGAGCAGCAAAAGCACCAGCGGCGGCACGGCTGCCCTGGGCTGCCTGGTTAPAGATTATTTCCCGGAACCAGTCACCGTGAGCTGGAACAGCGGGGCGCTGACCAGCGGCGTGCATACCTTTCCGGCGGTGCTGCAAAGCAGCGGCCTGTATAGCCTGAGCAGCGTTGTGACCGTGCCGAGCAGCAGCTTAGGCACTCAGACCTATATTTGCAACGTGAACCATAAACCGAGCAACACCAAAGTGGATAAAAAAGT GGAACCGAAAAGC X (X can be TGC,GAATTC orTGCGAATTC) PN encoding 144:GATATCGCACTGACCCAGCCAGCTTCAGTGAG SEQID NO: 140 CGGCTCACCAGGTCAGAGCATTACCATCTCGTGTACGGGTACTAGCAGCGATGTTGTTGGTGTTTATAATTTTGTGTCTTGGTACCAGCAGCATCCCGGGAAGGCGCCGAAACTTATGATTTATTATGTTGATAATCGTCCCTCAGGCGTGAGCAACCGTTTTAGCGGATCCAAAAGCGGCAACACCGCGAGCCTGACCATTAGCGGCCTGCAAGCGGAAGACGAAGCGGATTATTATTGCCAGTCTTTTGATGGTTTTGGTATTGATATGGTGTTTGGCGGCGGCACGAAGTTAACCGTTCTTGGCCAGCCGAAAGCCGCACCGAGTGTGACGCTGTTTCCGCCGAGCAGCGAAGAATTGCAGGCGAACAAAGCGACCCTGGTGTGCCTGATTAGCGACTTTTATCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAGGGGAGCACCGTGGAAAAAACCGTTGCGCCGAC TGAGX (X can be TGCAGC orGCC)Antibody 6755 CDRH1 145:SYWIA CDRH2 146:IIYPGDSDTNYSPSFQG CDRH3147:SKYGSFDY CDRL1 148:TGTSSDVGGYNYVS CDRL2 149:NVNSRPS CDRL3150:QSYDDGQDNEV VH 151:QVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIAWVRQMPGKGLEWMGIIYPGDSDTNYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARSKYGSF DYWGQGTLVTVSS VL152:DIALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYNVNSRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCQSYODGQDNEVFGG GTKLTVL Heavy chain153:QVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIAWVRQMPGKGLEWMGIIYPGDSDTNYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARSKYGSFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KKVEPKSX (X can be G, EF or CEF)Light chain 154:DIALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYNVNSRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCQSYODGQDNEVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVA PTEX (X can be CS or A) PN encoding155:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 151GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTTCTTATTGGATTGCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATCTATCCGGGTGATAGCGATACCAATTATTCTCCGAGCTTTCAGGGCCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTCTAAGTATGGTTCTTTTGATTATTG GGGCCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding 156:GATATCGCACTGACCCAGCCAGCTTCAGTGAG SEQ ID NO: 152CGGCTCACCAGGTCAGAGCA1TACCATCTCGTGTACGGGTACTAGCAGCGATGTTGGTGGTTATAATTATGTGTCTTGGTACCAGCAGCATCCCGGGAAGGCGCCGAAACTTATGATTTATAATGTTAATTCTCGTCCCTCAGGCGTGAGCAACCGTTTTAGCGGATCCAAAAGCGGCAACACCGCGAGCCTGACCATTAGCGGCCTGCAAGCGGAAGACGAAGCGGATTATTATTGCCAGTCTTATGATGATGGTCAGGATAATGAGGTGTTTGGCGGCGGCACGAA GTTAACCGTTCTT PN encoding157:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 153GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTTCTTATTGGATTGCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATCTATCCGGGTGATAGCGATACCAATTATTCTCCGAGCTTTCAGGGCCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTCTAAGTATGGTTCTTTTGATTATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCGTCGACCAAAGGTCCAAGCGTGTTTCCGCTGGCTCCGAGCAGCAAAGCACCAGCGGCGGCACGGCTGCCCTGGGCTGCCTGGTTAAAGATTATTTCCCGGAACCAGTCACCGTGAGCTGGAACAGCGGGGCGCTGACCAGCGGCGTGCATACCTTTCCGGCGGTGCTGCAAAGCAGCGGCCTGTATAGCCTGAGCAGCGTTGTGACCGTGCCGAGCAGCAGCTTAGGCACTCAGACCTATATTTGCAACGTGAACCATAAACCGAGCAACACCAAAGTGGATAAAAAAGTG GAACCGAAAAGC X (X can be TGC,GAATTC or TGCGAATTC) PN encoding 158:GATATCGCACTGACCCAGCCAGCTTCAGTGAGSEQ ID NO: 154 CGGCTCACCAGGTCAGAGCATTACCATCTCGTGTACGGGTACTAGCAGCGATGTTGGTGGTTATAATTATGTGTCTTGGTACCAGCAGCATCCCGGGAAGGCGCCGAAACTTATGATTTATAATGTTAATTCTCGTCCCTCAGGCGTGAGCAACCGTTTTAGCGGATCCAAAAGCGGCAACACCGCGAGCCTGACCATTAGCGGCCTGCAAGCGGAAGACGAAGCGGATTATTATTGCCAGTCTTATGATGATGGTCAGGATAATGAGGTGTTTGGCGGCGGCACGAAGTTAACCGTTCTTGGCCAGCCGAAAGCCGCACCGAGTGTGACGCTGTTTCCGCCGAGCAGCGAAGAATTGCAGGCGAACAAAGCGACCCTGGTGTGCCTGATTAGCGACTTTTATCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAGGGGAGCACCGTGGAAAAAACCGTTGCGCCGACTGA GX (X can be TGCAGC or GCC)Antibody 6757 CDRH1 159: SYAMH CDRH2 160: AISSSGSSTYYADSVKG CDRH3161:ESWFLDL CDRL1 162:RASQSISNWLA CDRL2 163:LASSLQS CDRL3 164:QQYYOFSDTVH 165:QVQLVESGGGLVQPGGSLRLSCAASGFTFTSYAMHWVRQAPGKGLEVWSAISSSGSSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARESWFLD LWGQGTLVTVSS VL166:DIQMTQSPSSLSASVGDRVTITCRASQSISNWLAWYQQKPGKAPKLLIYLASSLQSGVPSRFSGSGSGTDFTLTISSLQPEOFAVYYCQQYYDFSDTFGQGTKV EIK Heavy chain167:QVQLVESGGGLVQPGGSLRLSCAASGFTFTSYAMHWVRQAPGKGLEWVSAISSSGSSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARESWFLDLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKK VEPKSX (X can be C, EF or CEF)Light chain 168:DIQMTQSPSSLSASVGORVTITCRASQSISNWLAWYQQKPGKAPKLLIYLASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFAVYYCQQYYDFSDTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRG EX (X can be C or A) PN encoding169:CAGGTGCAATTGGTGGAAAGCGGCGGCGGCCT SEQ ID NO: 165GGTGCAACCGGGCGGCAGCCTGCGTCTGAGCTGCGCGGCCTCCGGATTTACCTTTACTTCTTATGCTATGCATTGGGTGCGCCAAGCCCCTGGGAAGGGTCTCGAGTGGGTGAGCGCTATCTCTTCTTCTGGTAGCTCTACCTATTATGCGGATAGCGTGAAAGGCCGTTTTACCATTTCACGTGATAATTCGAAAAACACCCTGTATCTGCAAATGAACAGCCTGCGTGCGGAAGATACGGCCGTGTATTATTGCGCGCGTGAGTCTTGGTTTCTTGATCTTTGGGG CCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding 170:GATATCCAGATGACCCAGAGCCCGTCTAGCCT SEQ ID NO: 166GAGCGCGAGCGTGGGTGATCGTGTGACCATTACCTGCAGAGCGAGCCAGTCTATTTCTAATTGGCTGGCTTGGTACCAGCAGAAACCAGGTAAAGCACCGAAACTATTAATTTATCTTGCTTCTTCTTTGCAAAGCGGGGTCCCGTCCCGTTTTAGCGGCTCTGGATCCGGCACTGATTTTACCCTGACCATTAGCAGCCTGCAACCTGAAGACTTTGCGGTTTATTATTGCCAGCAGTATTATGATTTTTCTGATACCTTTGGCCAGGGTACGAAAGTTGAAATTAA A PN encoding171:CAGGTGCAATTGGTGGAAAGCGGCGGCGGCCT SEQ ID NO: 167GGTGCAACCGGGCGGCAGCCTGCGTCTGAGCTGCGCGGCCTCCGGATTTACCTTTACTTCTTATGCTATGCATTGGGTGCGCCAAGCCCCTGGGAAGGGTCTCGAGTGGGTGAGCGCTATCTCTTCTTCTGGTAGCTCTACCTATTATGCGGATAGCGTGAAAGGCCGT1TTACCATTTCACGTGATAATTCGAAAAACACCCTGTATCTGCAAATGAACAGCCTGCGTGCGGAAGATACGGCCGTGTATTATTGCGCGCGTGAGTCTTGGTTTCTTGATCTTTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCGTCGACCAAAGGTCCAAGCGTGTTTCCGCTGGCTCCGAGCAGCAAAAGCACCAGCGGCGGCACGGCTGCCCTGGGCTGCCTGGTTAAAGATTATTTCCCGGAACCAGTCACCGTGAGCTGGAACAGCGGGGCGCTGACCAGCGGCGTGCATACCTTTCCGGCGGTGCTGCAAAGCAGCGGCCTGTATAGCCTGAGCAGCGTTGTGACCGTGCCGAGCAGCAGCTTAGGCACTCAGACCTATATTTGCAACGTGAACCATAAACCGAGCAACACCAAAGTGGATAAAAAAGTGGA ACCGAAAAGCX (X can be TGC, GAATTCor TGCGAATTC) PN encoding 172:GATATCCAGATGACCCAGAGCCCGTCTAGCCT SEQ IDNO: 168 GAGCGCGAGCGTGGGTGATCGTGTGACCATTACCTGCAGAGCGAGCCAGTCTATTTCTAATTGGCTGGCTTGGTACCAGCAGAAACCAGGTAAAGCACCGAAACTATTAATTTATCTTGCTTCTTCTTTGCAAAGCGGGGTCCCGTCCCGTTTTAGCGGCTCTGGATCCGGCACTGATTTTACCCTGACCATTAGCAGCCTGCAACCTGAAGACTTTGCGGTTTATTATTGCCAGCAGTATTATGATTTTTCTGATACCTTTGGCCAGGGTACGAAAGTTGAAATTAAACGTACGGTGGCTGCTCCGAGCGTGTTTATTTTTCCGCCGAGCGATGAACAACTGAAAAGCGGCACGGCGAGCGTGGTGTGCCTGCTGAACAACTTTTATCCGCGTGAAGCGAAAGTTCAGTGGAAAGTAGACAACGCGCTGCAAAGCGGCAACAGCCAGGAAAGCGTGACCGAACAGGATAGCAAAGATAGCACCTATTCTCTGAGCAGCACCCTGACCCTGAGCAAAGCGGATTATGAAAAACATAAAGTGTATGCGTGCGAAGTGACCCATCAAGGTCTGAGCAG CCCGGTGACTAAATCTTTTAATCGTGGCGAGX (Xcan be TGC or GCC) Antibody 6763 CDRH1 173:NYGMH CDRH2174:VSYAGSFTNYADSVKG CDRH3 175:SWLGYPDIFDY CDRL1 176:TGTSSDVGGYNYVSCDRL2 177:DVNNRPS CDRL3 178:SSYDKFQTV VH179:QVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGMHWVRQAPGKGLEWVSVSYAGSFTNYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSWLFGYP DIFDYWGQGTLVTVSS VL180:DIALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYDVNNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDKFQTVFGGGT KLTVL Heavy chain181:QVQLVESGGGLVQPGGSLRLSCMSGFTFSNYGMHWVRQAPGKGLEWISVSYAGSFTNYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSWLFGYPDIFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKV DKKVEPKSX (X can be C, EF or CEF)Light chain 182:DIALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYDVNNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDKFQTVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPT EX (X can be CS or A) PN encoding183:CAGGTGCAATTGGTGGAAAGCGGCGGCGGCCT SEQ ID NO: 179GGTGCAACCGGGCGGCAGCCTGCGTCTGAGCTGCGCGGCCTCCGGATTTACCTTTTCTAATTATGGTATGCATTGGGTGCGCCAAGCCCCTGGGAAGGGTCTCGAGTGGGTGAGCGTTTCTTATGCTGGTAGCTTTACCAATTATGCGGATAGCGTGAAAGGCCGTTTTACCATTTCACGTGATAATTCGAAAAACACCCTGTATCTGCAAATGAACAGCCTGCGTGCGGAAGATACGGCCGTGTATTATTGCGCGCGTTCTTGGCTTTTTGGTTATCCTGATATTTTTGATTATTGGGGCCAAGGCACCCTGGTGACGGTTAG CTCA PN encoding184:GATATCGCACTGACCCAGCCAGCTTCAGTGAG SEQ ID NO: 180CGGCTCACCAGGTCAGAGCA1TACCATCTCGTGTACGGGTACTAGCAGCGATGTTGGTGGTTATAATTATGTGTCTTGGTACCAGCAGCATCCCGGGAAGGCGCCGAAACTTATGATTTATGATGTTAATAATCGTCCCTCAGGCGTGAGCAACCGTTTTAGCGGATCCAAAAGCGGCAACACCGCGAGCCTGACCATTAGCGGCCTGCAAGCGGAAGACGAAGCGGATTATTATTGCTCTTCTTATGATAAGTTTCAGACTGTGTTTGGCGGCGGCACGAAGTTAAC CGTTCTT PN encoding185:CAGGTGCAATTGGTGGAAAGCGGCGGCGGCCT SEQ ID NO: 181GGTGCAACCGGGCGGCAGCCTGCGTCTGAGCTGCGCGGCCTCCGGATTTACCTTTTCTAATTATGGTATGCATTGGGTGCGCCAAGCCCCTGGGAAGGGTCTCGAGTGGGTGAGCGTTTCTTATGCTGGTAGCTTTACCAATTATGCGGATAGCGTGAAAGGCCGTTTTACCATTTCACGTGATAATTCGAAAAACACCCTGTATCTGCAAATGAACAGCCTGCGTGCGGAAGATACGGCCGTGTATTATTGCGCGCGTTCTTGGCTTTTTGGTTATCCTGATATTTTTGATTATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCGTCGACCAAAGGTCCAAGCGTGTTTCCGCTGGCTCCGAGCAGCAAAAGCACCAGCGGCGGCACGGCTGCCCTGGGCTGCCTGGTTAAAGATTATTTCCCGGAACCAGTCACCGTGAGCTGGAACAGCGGGGCGCTGACCAGCGGCGTGCATACCTTTCCGGCGGTGCTGCAAAGCAGCGGCCTGTATAGCCTGAGCAGCGTTGTGACCGTGCCGAGCAGCAGCTTAGGCACTCAGACCTATATTTGCAACGTGAACCATAAACCGAGCAACACCAAAGTGGA TAAAAAAGTGGAACCGAAAAGCX (X can beTGC, GAATTC or TGCGAATTC) PN encoding186:GATATCGCACTGACCCAGCCAGCTTCAGTGAG SEQ ID NO: 182CGGCTCACCAGGTCAGAGCA1TACCATCTCGTGTACGGGTACTAGCAGCGATGTTGGTGGTTATAATTATGTGTCTTGGTACCAGCAGCATCCCGGGAAGGCGCCGAAACTTATGATTTATGATGTTAATAATCGTCCCTCAGGCGTGAGCAACCGTTTTAGCGGATCCAMAGCGGCAACACCGCGAGCCTGACCATTAGCGGCCTGCAAGCGGAAGACGAAGCGGATTATTATTGCTCTTCTTATGATAAGTTTCAGACTGTGTTTGGCGGCGGCACGAAGTTAACCGTTCTTGGCCAGCCGAAAGCCGCACCGAGTGTGACGCTGTTTCCGCCGAGCAGCGAAGAATTGCAGGCGAACAAAGCGACCCTGGTGTGCCTGATTAGCGACTTTTATCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAGGGGAGC ACCGTGGAAAAAACCGTTGCGCCGACTGAGX (Xcan be TGCAGC or GCC) Antibody 7085 CDRH1 SEQ ID NO: 1 CDRH2 SEQ ID NO:2 CDRH3 SEQ ID NO: 3 CDRL1 SEQ ID NO: 4 CDRL2 SEQ ID NO: 5 CDRL3 SEQ IDNO: 6 VH 187:QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISVWRQAPGQGLEWMGGIGPFFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARDTPYFD YWGQGTLVTVSS VL188:DIELTQPPSVSVAPGQTARISCSGDSIPNYYVYWYQQKPGQAPVLVIYDDSNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQSFDSSLNAEVFGGGTK LTVL Heavy chain189:QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIGPFFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCAROTPYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK KVEPKSX (X can be G, EF or CEF)Light chain 190:DIELTQPPSVSVAPGQTARISCSGDSIPNYYVYWYQQKPGQAPVLVIYDDSNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQSFDSSLNAEVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTE X (X can be CS or A) PN encoding191:CAGGTGCAATTGGTTCAGTCTGGCGCGGAAGT SEQ ID NO: 187GAAAAAACCGGGCAGCAGCGTGAAAGTGAGCTGCAAAGCCTCCGGAGGCACTTTTTCTTCTTATGCCATTTCTTGGGTGCGCCAAGCCCCTGGGCAGGGTCTCGAGTG GATGGGCGGTATCGGTCCGTTTTTTGGCACTGCGA ATTACGCGCAGAAGTTTCAGGGCCGGGTGACCATTACCGCGGATGAAAGCACCAGCACCGCGTATATGGAACTGAGCAGCCTGCGTAGCGAAGATACGGCCGTGTATTATTGCGCGCGTGATACTCCTTATTTTGATTATTGGG GCCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding 192:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 188CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTATTCCTAATTATTATGTTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATGATGATTCTAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGTCTTTTGATTCTTCTCTTAATGCTGAGGTGTTTGGCGGCGGCACGAAGTTAACCGT TCTT PN encoding193:CAGGTGCAATTGGTTCAGTCTGGCGCGGAAGT SEQ ID NO: 189GAAAAAACCGGGCAGCAGCGTGAAAGTGAGCTGCAAAGCCTCCGGAGGCACTTTTTCTTCTTATGCCATTTCTTGGGTGCGCCAAGCCCCTGGGCAGGGTCTCGAGTGGATGGGCGGTATCGGTCCGTTTTTTGGCACTGCGAATTACGCGCAGAAGTTTCAGGGCCGGGTGACCATTACCGCGGATGAAAGCACCAGCACCGCGTATATGGAACTGAGCAGCCTGCGTAGCGAAGATACGGCCGTGTATTATTGCGCGCGTGATACTCCTTATTTTGATTATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCGTCGACCAAAGGTCCAAGCGTGTTTCCGCTGGCTCCGAGCAGCAAAAGCACCAGCGGCGGCACGGCTGCCCTGGGCTGCCTGGTTAAAGATTATTTCCCGGAACCAGTCACCGTGAGCTGGAACAGCGGGGCGCTGACCAGCGGCGTGCATACCTTTCCGGCGGTGCTGCAAAGCAGCGGCCTGTATAGCCTGAGCAGCGTTGTGACCGTGCCGAGCAGCAGCTTAGGCACTCAGACCTATATTTGCAACGTGAACCATAAACCGAGCAACACCAAAGTGGATAAAAAAGTGGA ACCGAAAAGCX (X can be TGC, GAATTCor TGCGAATTC) PN encoding 194:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ IDNO: 190 CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTATTCCTAATTATTATGTTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATGATGATTCTAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGTCTTTTGATTCTTCTCTTAATGCTGAGGTGTTTGGCGGCGGCACGAAGTTAACCGTTCTTGGCCAGCCGAAAGCCGCACCGAGTGTGACGCTGTTTCCGCCGAGCAGCGAAGAATTGCAGGCGAACAAAGCGACCCTGGTGTGCCTGATTAGCGACTTTTATCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAGGGGAGCAC CGTGGAAAAAACCGTTGCGCCGACTGAGX (Xcan be TGCAGC or GCC) Antibody 7087 CDRH1 195:SYYIS CDRH2196:GIIPIFGTANYAQKFQG CDRH3 197:GEIWHTHQPYKSGVYGAAY CDRL1198:RASQGISNWLN CDRL2 199:GTSSLQS CDRL3 200:QQLDSFPAT VH201:QTQLVQSGAEVKKPGSSVKVSCKASGGTFSSYYISWVRQAPGQGLEWMGGIPTFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGEIWHVH QPYKSGVYGAAYWGQGTLVTVSS VL202:DIQMTQSPSSLSASVGDRVTITCRASQGISNWLNWYQQKPGKAPKLLIYGTSSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQLDSFPATFGQGTKV EIK Heavy chain203:QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYYISWVRQAPGQGLEWMGGIFTFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGEIWHVHQPYKSGVYGAAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV NHKPSNTKVDKKVEPKSX(X can be C, EFor CEF) Light chain 204:DIQMTQSPSSLSASVGDRVTITCRASQGISNWLNWYQQKPGKAPKLLIYGTSSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQLDSFPATFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRG EX (X can be C or A) PN encoding205:CAGGTGCAATTGGTTCAGTCTGGCGCGGAAGT SEQ ID NO: 201GAAAAAACCGGGCAGCAGCGTGAAAGTGAGCTGCAAAGCCTCCGGAGGCACTTTTTCTTCTTATTATATTTCTTGGGTGCGCCAAGCCCCTGGGCAGGGTCTCGAGTGGATGGGCGGTATCATTCCGATTTTTGGCACTGCGAATTACGCGCAGAAGTTTCAGGGCCGGGTGACCATTACCGCGGATGAAAGCACCAGCACCGCGTATATGGAACTGAGCAGCCTGCGTAGCGAAGATACGGCCGTGTATTATTGCGCGCGTGGTGAGATTTGGCATGTTCATCAGCCTTATAAGTCTGGTGTTTATGGTGCTGCTTATTGGGG CCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding 206:GATATCCAGATGACCCAGAGCCCGTCTAGCCT SEQ ID NO: 202GAGCGCGAGCGTGGGTGATCGTGTGACCATTACCTGCAGAGCGAGCCAGGGTATTTCTAATTGGCTGAATTGGTACCAGCAGAAACCAGGTAAAGCACCGAAACTATTAATTTATGGTACTTCTTCTTTGCAAAGCGGGGTCCCGTCCCGTTTTAGCGGCTCTGGATCCGGCACTGATTTTACCCTGACCATTAGCAGCCTGCAACCTGAAGACTTTGCGACTTATTATTGCCAGCAGCTTGATTCTTTTCCTGCTACCTTTGGCCAGGGTACGAAAGTTGAAATTAA A PN encoding207:CAGGTGCAATTGGTTCAGTCTGGCGCGGAAGT SEQ ID NO: 203GAAAAAACCGGGCAGCAGCGTGAAAGTGAGCTGCAAAGCCTCCGGAGGCACTTTTTCTTCTTATTATATTTCTTGGGTGCGCCAAGCCCCTGGGCAGGGTCTCGAGTGGATGGGCGGTATCATTCCGATTTTTGGCACTGCGAATTACGCGCAGAAGTTTCAGGGCCGGGTGACCATTACCGCGGATGAAAGCACCAGCACCGCGTATATGGAACTGAGCAGCCTGCGTAGCGAAGATACGGCCGTGTATTATTGCGCGCGTGGTGAGATTTGGCATGTTCATCAGCCTTATAAGTCTGGTGTTTATGGTGCTGCTTATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCGTCGACCAAAGGTCCAAGCGTGTTTCCGCTGGCTCCGAGCAGCAAAAGCACCAGCGGCGGCACGGCTGCCCTGGGCTGCCTGGTTAAAGATTATTTCCCGGAACCAGTCACCGTGAGCTGGAACAGCGGGGCGCTGACCAGCGGCGTGCATACCTTTCCGGCGGTGCTGCAAAGCAGCGGCCTGTATAGCCTGAGCAGCGTTGTGACCGTGCCGAGCAGCAGCTTAGGCACTCAGACCTATATTTGCAACGTGAACCATAAACCGAGCAACACCAAAGTGGATAAAAAAGTGGA ACCGAAAAGCX (X can be TGC, GAATTCor TGCGAATTC) PN encoding 208:GATATCCAGATGACCCAGAGCCCGTCTAGCCT SEQ IDNO: 204 GAGCGCGAGCGTGGGTGATCGTGTGACCATTACCTGCAGAGCGAGCCAGGGTATTTCTAATTGGCTGAATTGGTACCAGCAGAAACCAGGTAAAGCACCGAAACTATTAATTTATGGTACTTCTTCTTTGCAAAGCGGGGTCCCGTCCCGTTTTAGCGGCTCTGGATCCGGCACTGATTTTACCCTGACCATTAGCAGCCTGCAACCTGAAGACTTTGCGACTTATTATTGCCAGCAGCTTGATTCTTTTCCTGCTACCTTTGGCCAGGGTACGAAAGTTGAAATTAAACGTACGGTGGCTGCTCCGAGCGTGTTTATTTTTCCGCCGAGCGATGAACAACTGAAAAGCGGCACGGCGAGCGTGGTGTGCCTGCTGAACAACTTTTATCCGCGTGAAGCGAAAGTTCAGTGGAAAGTAGACAACGCGCTGCAAAGCGGCAACAGCCAGGAAAGCGTGACCGAACAGGATAGCAAAGATAGCACCTATTCTCTGAGCAGCACCCTGACCCTGAGCAAAGCGGATTATGAAAAACATAAAGTGTATGCGTGCGAAGTGACCCATCAAGGTCTGAGCAG CCCGGTGACTAAATCTTTTAATCGTGGCGAGX (Xcan be TGC or GCC) Antibody 7091 CDRH1 SEQ ID NO: 61 CDRH2 SEQ ID NO: 77CDRH3 SEQ ID NO: 63 CDRL1 SEQ ID NO: 64 CDRL2 SEQ ID NO: 65 CDRL3209:QSWTDSPNTLV VH 210:QVQLVQSGAEVKKPGESLKISCKGSGYSFTSYYIGWIRQMPGKGLEWMGIIDPSDSHTTYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYMMRGF DHWGQGTLVTVSS VL211:DIELTQPPSVSVAPGQTARISCSGDSLGDYYAYWYQQKPGQAPVLVIYKDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQSWTDSPNTLVFGGGTK LTVL Heavy chain212:QVQLVQSGAEVKKPGESLKISCKGSGYSFTSYYIGWVRQMPGKGLEWMGIIDPSDSHTTYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYMMRGFDHWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDK KVEPKSX (X can be C, EF or CEF)Light chain 213:DIELTQPPSVSVAPGQTARISCSGDSLGDYYAYWYQQKPGQAPVLVIYKDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQSWTDSPNTLVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTE X (X can be CS or A) PN encoding214:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 210GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTTCTTATTATATTGGTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATCGATCCGTCTGATAGCCATACCACTTATTCTCCGAGCTTTCAGGGCCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATATGATGCGTGGTTTTGATCATTG GGGCCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding 215:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 211CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTCTTGGTGATTATTATGCTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGTCTTGGACTGATTCTCCTAATACTCTTGTGTTTGGCGGCGGCACGAAGTTAACCGT TCTT PN encoding216:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 212GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTTCTTATTATATTGGTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATCGATCCGTCTGATAGCCATACCACTTATTCTCCGAGCTTTCAGGGCCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATATGATGCGTGGTTTTGATCATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCGTCGACCAAAGGTCCAAGCGTGTTTCCGCTGGCTCCGAGCAGCAAAAGCACCAGCGGCGGCACGGCTGCCCTGGGCTGCCTGGTTAAAGATTATTTCCCGGAACCAGTCACCGTGAGCTGGAACAGCGGGGCGCTGACCAGCGGCGTGCATACCTTTCCGGCGGTGCTGCAAAGCAGCGGCCTGTATAGCCTGAGCAGCGTTGTGACCGTGCCGAGCAGCAGCTTAGGCACTCAGACCTATATTTGCAACGTGAACCATAAACCGAGCAACACCAAAGTGGATAAAAAAGT GGAACCGAAAAGCX (X can be TGC,GAATTC or TGCGAATTC) PN encoding 217:GATATCGAACTGACCCAGCCGCCTTCAGTGAGSEQ ID NO: 213 CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTCTTGGTGATTATTATGCTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGTCTTGGACTGATTCTCCTAATACTCTTGTGTTTGGCGGCGGCACGAAGTTAACCGTTCTTGGCCAGCCGAAAGCCGCACCGAGTGTGACGCTGTTTCCGCCGAGCAGCGAAGAATTGCAGGCGAACAAAGCGACCCTGGTGTGCCTGATTAGCGACTTTTATCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAGGGGAGCAC CGTGGAAAAAACCGTTGCGCCGACTGAGX (Xcan be TGCAGC or GCC) Antibody 7092 CDRH1 SEQ ID NO: 17 CDRH2 SEQ ID NO:49 CDRH3 SEQ ID NO: 19 CDRL1 SEQ ID NO: 20 CDRL2 SEQ ID NO: 21 CDRL3 SEQID NO: 22 VH 218:QVQLVQSGAEVKKPGESLKISCKGSGYSFTNYISVWRQMPGKGLEWMGIIDPDDSYTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYEYGGFD IWGQGTLVTVSS VL219:DIELTQPPSVSVAPGQTARISCSGDNIGNSYVHWYQQKPGQAPVLVIYKDNDRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCGTYDIESYVFGGGTKLT VL Heavy chain220:QVQLVQSGAEVKKPGESLKISCKGSGYSFTNYISVWRQMPGKGLEWMGIIDPDDSYTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYEYGGFDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKK VEPKSX (X can be C, EF or CEF)Light chain 221:DIELTQPPSVSVAPGQTARISCSGDNIGNSYVHWYQQKPGQAPVLVIYKDNDRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCGTYDIESYVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASS YLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTEX(X can be CS or A) PN encoding 222:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQID NO: 218 GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTAATTATATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATCGATCCGGATGATAGCTATACCCGTTATTCTCCGAGCTTTCAGGGACAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATGAGTATGGTGGTTTTGATATTTGGGG CCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding 223:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 219CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATAATATTGGTAATTCTTATGTTCATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATGATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCGGTACTTATGATATTGAGTCTTA TGTGTTTGGCGGCGGCACGAAGTTAACCGTTCTTPN encoding 224:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 220GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTAATTATATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATCGATCCGGATGATAGCTATACCCGTTATTCTCCGAGCTTTCAGGGACAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATGAGTATGGTGGTTTTGATATTTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCGTCGACCAAAGGTCCAAGCGTGTTTCCGCTGGCTCCGAGCAGCAAAAGCACCAGCGGCGGCACGGCTGCCCTGGGCTGCCTGGTTAAAGATTATTTCCCGGAACCAGTCACCGTGAGCTGGAACAGCGGGGCGCTGACCAGCGGCGTGCATACCTTTCCGGCGGTGCTGCAAAGCAGCGGCCTGTATAGCCTGAGCAGCGTTGTGACCGTGCCGAGCAGCAGCTTAGGCACTCAGACCTATATTTGCAACGTGAACCATAAACCGAGCAACACCAAAGTGGATAAAAAAGTGGA ACCGAAAAGCX (X can be TCC, GAATTCor TGCGAATTC) PN encoding 225:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ IDNO: 221 CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATAATATTGGTAATTCTTATGTTCATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATGATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCGGTACTTATGATATTGAGTCTTATGTGTTTGGCGGCGGCACGAAGTTAACCGTTCTTGGCCAGCCGAAAGCCGCACCGAGTGTGACGCTGTTTCCGCCGAGCAGCGAAGAATTGCAGGCGAACAAAGCGACCCTGGTGTGCCTGATTAGCGACTTTTATCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAGGGGAGCACCGTGGA AAAAACCGTTGCGCCGACTGAGX (X can beTGCAGC or GCC) Antibody 7093 CDRH1 SEQ ID NO: 33 CDRH2226:HIFSDDDKYYSTSLKT CDRH3 SEQ ID NO: 35 CDRL1 SEQ ID NO: 36 CDRL2 SEQID NO: 37 CDRL3 SEQ ID NO: 38 VH 227:QVQLKESGPALVKPTQTLTLTCTFSGFSLSTSGGGVSWIRQPPGKALEWLAHIFSDDDKYYSTSLKTRLTISKDTSKNQVVLTMTNMDPVDTATYYCARGPYDF DSWGQGTLVTVSS VL228:DIALTQPASVSGSPGQSITISCTGTSSDIGTYNYVSWYQQHPGKAPKLMIYDDSNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCQSYDSQSIVFGGGT KLTVL Heavy chain229:QVQLKESGPALVKPTQTLTLTCTFSGFSLSTSGGGVSWIRQPPGKALEWLAHIFSDDDKYYSTSLKTRLTISKDTSKNQVVLTMTNMDPVDTATYYCARGPYDFDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGDLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KKVEPKSX (X can be C, EF or CEF)Light chain 230:DIALTQPASVSGSPGQSITISCTGTSSDIGTYNYVSWYQQHPGKAPKLMIYDDSNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCQSYDSQSIVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPT EX (X can be CS orA) PN encoding231:CAGGTGCAATTGAAAGAAAGCGGCCCGGCCCT SEQ ID NO: 227GGTGAAACCGACCCAAACCCTGACCCTGACCTGTACCTTTTCCGGATTTAGCCTGTCTACTTCTGGTGGTGGTGTGTCTTGGATTCGCCAGCCGCCTGGGAAAGCCCTCGAGTGGCTGGCTCATATCTTTTCTGATGATGATAAGTATTATAGCACCAGCCTGAAAACGCGTCTGACCATTAGCAAAGATACTTCGAAAAATCAGGTGGTGCTGACTATGACCAACATGGACCCGGTGGATACGGCCACCTATTATTGCGCGCGTGGTCCTTATGGTTTTGATTCTTG GGGCCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding 232:GATATCGCACTGACCCAGCCAGCTTCAGTGAG SEQ ID NO: 228CGGCTCACCAGGTCAGAGCATTACCATCTCGTGTACGGGTACTAGCAGCGATATTGGTACTTATAATTATGTGTCTTGGTACCAGCAGCATCCCGGGAAGGCGCCGAAACTTATGATTTATGATGATTCTAATCGTCCCTCAGGCGTGAGCAACCGTTTTAGCGGATCCAAAAGCGGCAACACCGCGAGCCTGACCATTAGCGGCCTGCAAGCGGAAGACGAAGCGGATTATTATTGCCAGTCTTATGATTCTCAGTCTATTGTGTTTGGCGGCGGCACGAAGTTAAC CGTTCTT PN encoding233:CAGGTGCAATTGAAAGAAAGCGGCCCGGCCCT SEQ ID NO: 229GGTGAAACCGACCCAAACCCTGACCCTGACCTGTACCTTTTCCGGATTTAGCCTGTCTACTTCTGGTGGTGGTGTGTCTTGGATTCGCCAGCCGCCTGGGAAAGCCCTCGAGTGGCTGGCTCATATCTTTTCTGATGATGATAAGTATTATAGCACCAGCCTGAAAACGCGTCTGACCATTAGCAAAGATACTTCGAAAAATCAGGTGGTGCTGACTATGACCAACATGGACCCGGTGGATACGGCCACCTATTATTGCGCGCGTGGTCCTTATGGTTTTGATTCTTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCGTCGACCAAAGGTCCAAGCGTGTTTCCGCTGGCTCCGAGCAGCAAAAGCACCAGCGGCGGCACGGCTGCCCTGGGCTGCCTGGTTAAAGATTATTTCCCGGAACCAGTCACCGTGAGCTGGAACAGCGGGGCGCTGACCAGCGGCGTGCATACCTTTCCGGCGGTGCTGCAAAGCAGCGGCCTGTATAGCCTGAGCAGCGTTGTGACCGTGCCGAGCAGCAGCTTAGGCACTCAGACCTATATTTGCAACGTGAACCATAAACCGAGCAACACCAAAGTGGATAAAAAAGT GGAACCGAAAAGCX (X can be TGC,GAATTC or TGCGAATTC) PN encoding 234:GATATCGCACTGACCCAGCCAGCTTCAGTGAGSEQ ID NO: 230 CGGCTCACCAGGTCAGAGCATTACCATCTCGTGTACGGGTACTAGCAGCGATATTGGTACTTATAATTATGTGTCTTGGTACCAGCAGCATCCCGGGAAGGCGCCGAAACTTATGATTTATGATGATTCTAATCGTCCCTCAGGCGTGAGCAACCGTTTTAGCGGATCCAAAAGCGGCAACACCGCGAGCCTGACCATTAGCGGCCTGCAAGCGGAAGACGAAGCGGATTATTATTGCCAGTCTTATGATTCTCAGTCTATTGTGTTTGGCGGCGGCACGAAGTTAACCGTTCTTGGCCAGCCGAAAGCCGCACCGAGTGTGACGCTGTTTCCGCCGAGCAGCGAAGAATTGCAGGCGAACAAAGCGACCCTGGTGTGCCTGATTAGCGACTTTTATCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAGGGGAG CACCGTGGAAAAAACCGTTGCGCCGACTGAGX (Xcan be TGCAGC or GCC) Antibody 7094 CDRH1 235:TSGMSVG CDRH2236:LIDWOEDKSYSTSLKT CDRH3 237:YNWYNPPGFDN CDRL1 238:SGSSSNIGSNYVS CDRL2239:RNDKRPS CDRL3 240:QSADSSSMV VH 241:QVQLKESGPALVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQPPGKALEWLALIDWDEOKSYSTSLKTRLTISKDTSKNQVVLTMTNMDPVDTATYYCARYNWYN PPGFDNWGQGTLVTVSS VL242:DIVLTQPPSVSGAPGQRVTISCSGSSSNIGSNYVSWYQQLPGTAPKLLIYRNDKRPSGVPDRFSGSKSGTSASLAITGLQSEDEADYYCQSADSSSMVFGGGTK LTVL Heavy chain243:QVQLKESGPALVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQPPGKALEWLALIDWDEOKSYSTSLKTRLTISKDTSKNQVVLTMTNMDPVDTATYYCARYNWYNPPGFDNWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN TKVDKKVEPKSX (X can be C, EF orCEF) Light chain 244:DIVLTQPPSVSGAPGQRVTISCSGSSSNIGSNYVSWYQQLPGTAPKLLIYRNDKRPSGVPDRFSGSKSGTSASLAITGLQSEDEADYYCQSADSSSMVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTE X (X can be CS orA) PN encoding245:CAGGTGCAATTGAAAGAAAGCGGCCCGGCCCT SEQ ID NO: 241GGTGAAACCGACCCAAACCCTGACCCTGACCTGTACCTTTTCCGGATTTAGCCTGTCTACTTCTGGTATGTCTGTGGGTTGGATTCGCCAGCCGCCTGGGAAAGCCCTCGAGTGGCTGGCTCTTATCGATTGGGATGAGGATAAGTCTTATAGCACCAGCCTGAAAACGCGTCTGACCATTAGCAAAGATACTTCGAAAAATCAGGTGGTGCTGACTATGACCAACATGGACCCGGTGGATACGGCCACCTATTATTGCGCGCGTTATAATTGGTATAATCCTCCTGGTTTTGATAATTGGGGCCAAGGCACCCTGGTGACGGT TAGCTCA PN encoding246:GATATCGTGCTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 242TGGCGCACCAGGTCAGCGTGTGACCATCTCGTGTAGCGGCAGCAGCAGCAACATTGGTTCTAATTATGTGTCTTGGTACCAGCAGTTGCCCGGGACGGCGCCGAAACTTCTGATTTATCGTAATGATAAGCGTCCCTCAGGCGTGCCGGATCGTTTTAGCGGATCCAAAAGCGGCACCAGCGCGAGCCTTGCGATTACGGGCCTGCAAAGCGAAGACGAAGCGGATTATTATTGCCAGTCTGCTGATTCTTCTTCTATGGTGTTTGGCGGCGGCACGAAGTTAACCGT TCTT PN encoding247:CAGGTGCAATTGAAAGAAAGCGGCCCGGCCCT SEQ ID NO: 243GGTGAAACCGACCCAAACCCTGACCCTGACCTGTACCTTTTCCGGATTTAGCCTGTCTACTTCTGGTATGTCTGTGGGTTGGATTCGCCAGCCGCCTGGGAAAGCCCTCGAGTGGCTGGCTCTTATCGATTGGGATGAGGATAAGTCTTATAGCACCAGCCTGAAAACGCGTCTGACCATTAGCAAAGATACTTCGAAAAATCAGGTGGTGCTGACTATGACCAACATGGACCCGGTGGATACGGCCACCTATTATTGCGCGCGTTATAATTGGTATAATCCTCCTGGTTTTGATAATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCGTCGACCAAAGGTCCAAGCGTGTTTCCGCTGGCTCCGAGCAGCAAAAGCACCAGCGGCGGCACGGCTGCCCTGGGCTGCCTGGTTAAAGATTATTTCCCGGAACCAGTCACCGTGAGCTGGAACAGCGGGGCGCTGACCAGCGGCGTGCATACCTTTCCGGCGGTGCTGCAAAGCAGCGGCCTGTATAGCCTGAGCAGCGTTGTGACCGTGCCGAGCAGCAGCTTAGGCACTCAGACCTATATTTGCAACGTGAACCATAAACCGAGCAACACCAAAGT GGATAAAAAAGTGGAACCGAAAAGCX (X canbe TGC, GAATTC or TGCGAATTC) PN encoding248:GATATCGTGCTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 244TGGCGCACCAGGTCAGCGTGTGACCATCTCGTGTAGCGGCAGCAGCAGCAACATTGGTTCTAATTATGTGTCTTGGTACCAGCAGTTGCCCGGGACGGCGCCGAAACTTCTGATTTATCGTAATGATAAGCGTCCCTCAGGCGTGCCGGATCGTTTTAGCGGATCCAAAAGCGGCACCAGCGCGAGCCTTGCGATTACGGGCCTGCAAAGCGAAGACGAAGCGGATTATTATTGCCAGTCTGCTGATTCTTCTTCTATGGTGTTTGGCGGCGGCACGAAGTTAACCGTTCTTGGCCAGCCGAAAGCCGCACCGAGTGTGACGCTGTTTCCGCCGAGCAGCGAAGAATTGCAGGCGAACAAAGCGACCCTGGTGTGCCTGATTAGCGACTTTTATCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAGGGGAGCAC CGTGGAAAAAACCGTTGCGCCGACTGAGX (Xcan be TGCAGC or GCC) Antibody 7821 CDRH1 SEQ ID NO: 1 CDRH2 SEQ ID NO:119 CDRH3 SEQ ID NO: 3 CDRL1 SEQ ID NO: 4 CDRL2 SEQ ID NO: 5 CDRL3 SEQID NO: 6 VH SEQ ID NO: 121 VL SEQ ID NO: 188 Heavy chain249:QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGNIGPFFGIANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARDTPYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK KVEPKSX (X can be C, EF or CEF)Light chain SEQ ID NO: 190 PN encoding SEQ ID NO: 125 SEQ ID NO: 121 PNencoding SEQ ID NO: 192 SEQ ID NO: 188 PN encoding250:CAGGTGCAATTGGTTCAGTCTGGCGCGGAAGT SEQ ID NO: 249GAAAAAACCGGGCAGCAGCGTGAAAGTGAGCTGCAAAGCCTCCGGAGGCACTTTTTCTTCTTATGCCATTTCTTGGGTGCGCCAAGCCCCTGGGCAGGGTCTCGAGTGGATGGGCAATATCGGTCCGTTTTTTGGCATTGCGAATTACGCGCAGAAGTTTCAGGGCCGGGTGACCATTACCGCGGATGAAAGCACCAGCACCGCGTATATGGAACTGAGCAGCCTGCGTAGCGAAGATACGGCCGTGTATTATTGCGCGCGTGATACTCCTTATTTTGATTATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCGTCGACCAAAGGTCCAAGCGTGTTTCCGCTGGCTCCGAGCAGCAAAAGCACCAGCGGCGGCACGGCTGCCCTGGGCTGCCTGGTTAAAGATTATTTCCCGGAACCAGTCACCGTGAGCTGGAACAGCGGGGCGCTGACCAGCGGCGTGCATACCTTTCCGGCGGTGCTGCAAAGCAGCGGCCTGTATAGCCTGAGCAGCGTTGTGACCGTGCCGAGCAGCAGCTTAGGCACTCAGACCTATATTTGCAACGTGAACCATAAACCGAGCAACACCAAAGTGGATAAAAAAGTGGA ACCGAAAAGC X (X can be TGC, GAATTCor TGCGAATTC) PN encoding SEQ ID NO: 194 SEQ ID NO: 190 Antibody 7865CDRH1 SEQ ID NO: 1 CDRH2 SEQ ID NO: 2 CDRH3 SEQ ID NO: 3 CDRL1 SEQ IDNO: 4 CDRL2 SEQ ID NO: 5 CDRL3 SEQ ID NO: 120 VH SEQ ID NO: 187 VL SEQID NO: 122 Heavy chain SEQ ID NO: 189 Light chain251:DIELTQPPSTSTAPGQTARISCSGDSIPNYYTYWYQQKPGQAPVLVIYDDSNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQTYDDGSTAEVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTE X (X can be CS or A) PN encodingSEQ ID NO: 191 SEQ ID NO: 187 PN encoding SEQ ID NO: 126 SEQ ID NO: 122PN encoding SEQ ID NO: 193 SEQ ID NO: 189 PN encoding252:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 251CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTATTCCTAATTATTATGTTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATGATGATTCTAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGACTTATGATGATGGTTCTACTGCTGAGGTGTTTGGCGGCGGCACGAAGTTAACCGTTCTTGGCCAGCCGAAAGCCGCACCGAGTGTGACGCTGTTTCCGCCGAGCAGCGAAGAATTGCAGGCGAACAAAGCGACCCTGGTGTGCCTGATTAGCGACTTTTATCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAGGGGAGCAC CGTGGAAAAAACCGTTGCGCCGACTGAGX (Xcan be TGCAGC or GCC) Antibody 7829 CDRH1 SEQ ID NO: 61 CDRH2 SEQ ID NO:62 CDRH3 SEQ ID NO: 63 CDRL1 SEQ ID NO: 64 CDRL2 SEQ ID NO: 65 CDRL3 SEQID NO: 209 VH 253:QTQLTQSGAETKKPGESLKISCKGSGYSFTSYYIGWIRQMPGKGLEWMGIIDPTDSQTAYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYMMRGF DHWGQGTLVTVSS VL SEQ ID NO: 211Heavy chain 254:QVQLVQSGAEVKKPGESLKISCKGSGYSFTSYYIGWVRQMPGKGLEWMGIIDPTDSQTAYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYMMRGFDHWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDK KVEP KSX (X can be C, EF or CEF)Light chain SEQ ID NO: 213 PN encoding255:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 253GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTTCTTATTATATTGGTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTACTGATTCTCAGACTGCTTATTCTCCTTCTTTTCAGGGTCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATATGATGCGTGGTTTTGATCATTG GGGCCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding SEQ ID NO: 215 SEQ ID NO :211 PN encoding256:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 254GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTTCTTATTATATTGGTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTACTGATTCTCAGACTGCTTATTCTCCTTCTTTTCAGGGTCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATATGATGCGTGGTTTTGATCATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCGTCGACCAAAGGTCCAAGCGTGTTTCCGCTGGCTCCGAGCAGCAAAAGCACCAGCGGCGGCACGGCTGCCCTGGGCTGCCTGGTTAAAGATTATTTCCCGGAACCAGTCACCGTGAGCTGGAACAGCGGGGCGCTGACCAGCGGCGTGCATACCTTTCCGGCGGTGCTGCAAAGCAGCGGCCTGTATAGCCTGAGCAGCGTTGTGACCGTGCCGAGCAGCAGCTTAGGCACTCAGACCTATATTTGCAACGTGAACCATAAACCGAGCAACACCAAAGTGGATAAAAAAGT GGAACCGAAAAGCX (X can be TGC,GAATTC or TGCGAATTC) PN encoding SEQ ID NO: 217 SEQ ID NO: 213 Antibody7830 CDRH1 SEQ ID NO: 61 CDRH2 SEQ ID NO: 95 CDRH3 SEQ ID NO: 63 CDRL1SEQ ID NO: 64 CDRL2 SEQ ID NO: 65 CDRL3 SEQ ID NO: 209 VH257:QVQLVQSGAEVKKPGESLKISCKGSGYSFTSYYIGWIRQMPGKGLEWMGIIDPTDSYTVYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYMMRGF DHWGQGTLVTVSS VL SEQ ID NO: 211Heavy chain 258:QVQLVQSGAEVKKPGESLKISCKGSGYSFTSYYIGWVRQMPGKGLEWMGIIDPTDSYTVYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYMMRGFDHWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDK KVEPKSX (X can be C, EF or CEF)Light chain SEQ ID NO: 213 PN encoding259:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 257GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTTCTTATTATATTGGTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTACTGATTCTTATACTGTTTATTCTCCTTCTTTTCAGGGTCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATATGATGCGTGGTTTTGATCATTG GGGCCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding SEQ ID NO: 215 SEQ ID NO :211 PN encoding260:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 258GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTTCTTATTATATTGGTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTACTGATTCTTATACTGTTTATTCTCCTTCTTTTCAGGGTCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATATGATGCGTGGTTTTGATCATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCGTCGACCAAAGGTCCAAGCGTGTTTCCGCTGGCTCCGAGCAGCAAAAGCACCAGCGGCGGCACGGCTGCCCTGGGCTGCCTGGTTAAAGATTATTTCCCGGAACCAGTCACCGTGAGCTGGAACAGCGGGGCGCTGACCAGCGGCGTGCATACCTTTCCGGCGGTGCTGCAAAGCAGCGGCCTGTATAGCCTGAGCAGCGTTGTGACCGTGCCGAGCAGCAGCTTAGGCACTCAGACCTATATTTGCAACGTGAACCATAAACCGAGCAACACCAAAGTGGATAAAAAAGT GGAACCGAAAAGCX (X can be TGC,GAATTC or TGCGAATTC) PN encoding SEQ ID NO: 217 SEQ ID NO: 213 Antibody7871 CDRH1 SEQ ID NO: 61 CDRH2 SEQ ID NO: 77 CDRH3 SEQ ID NO: 63 CDRL1SEQ ID NO: 64 CDRL2 SEQ ID NO: 65 CDRL3 SEQ ID NO: 66 VH SEQ ID NO: 210VL 261:DIELTQPPSVSVAPGQTARISCSGDSLGDYYAYWYQQKPGQAPVLVIYKDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQTWDTGESGVFGGGTKL TVL Heavy chain SEQ ID NO: 212Light chain 262:DIELTQPPSVSVAPGQTARISCSGDSLGDYYAYWYQQKPGQAPVLVIYKDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQTWDTGESGVFGGGTKLTVLGQPKMPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASS YLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTEX(X can be CS orA) PN encoding SEQ ID NO: 214 SEQ ID NO: 210 PN encoding263:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 261CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTCTTGGTGATTATTATGCTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGACTTGGGATACTGGTGAGTCTGGTGTGTTTGGCGGCGGCACGAAGTTAACCGTTCT T PN encoding SEQ ID NO: 216 SEQ IDNO: 212 PN encoding 264:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 262CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTCTTGGTGATTATTATGCTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGACTTGGGATACTGGTGAGTCTGGTGTGTTTGGCGGCGGCACGAAGTTAACCGTTCTTGGCCAGCCGAAAGCCGCACCGAGTGTGACGCTGTTTCCGCCGAGCAGCGAAGAATTGCAGGCGAACAAAGCGACCCTGGTGTGCCTGATTAGCGACTTTTATCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAGGGGAGCACCGT GGAAAAAACCGTTGCGCCGACTGAGX (X canbe TGCAGC or GCC) Antibody 7872 CDRH1 SEQ ID NO: 61 CDRH2 SEQ ID NO: 77CDRH3 SEQ ID NO: 63 CDRL1 SEQ ID NO: 64 CDRL2 SEQ ID NO: 65 CDRL3 SEQ IDNO: 78 VH SEQ ID NO: 210 VL 265:DIELTQPPSVSVAPGQTARISCSGDSLGDYYAYWYQQKPGQAPVLVIYKDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQTWDILPHGLVFGGGTK LTVL Heavy chain SEQ ID NO: 212Light chain 266:DIELTQPPSVSVAPGQTARISCSGDSLGDYYAYWYQQKPGQAPVLVIYKDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQTWDILPHGLVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTE X (X can be CS orA) PN encoding SEQID NO: 214 SEQ ID NO: 210 PN encoding267:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 265CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTCTTGGTGATTATTATGCTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGACTTGGGATATTCTTCCTCATGGTCTTGTGTTTGGCGGCGGCACGAAGTTAACCGT TCTT PN encoding SEQ ID NO: 216 SEQID NO :212 PN encoding 268:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO:266 CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTCTTGGTGATTATTATGCTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGACTTGGGATATTCTTCCTCATGGTCTTGTGTTTGGCGGCGGCACGAAGTTAACCGTTCTTGGCCAGCCGAAAGCCGCACCGAGTGTGACGCTGTTTCCGCCGAGCAGCGAAGAATTGCAGGCGAACAAAGCGACCCTGGTGTGCCTGATTAGCGACTTTTATCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAGGGGAGCAC CGTGGAAAAAACCGTTGCGCCGACTGAGX (Xcan be TGCAGC or GCC) Antibody 7873 CDRH1 SEQ ID NO: 61 CDRH2 SEQ ID NO:77 CDRH3 SEQ ID NO: 63 CDRL1 SEQ ID NO: 64 CDRL2 SEQ ID NO: 65 CDRL3 SEQID NO: 89 VH SEQ ID NO: 210 VL 269:DIELTQPPSVSVAPGQTARISCSGDSLGDYYAYWYQQKPGQAPVLVIYKDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQAWTDSPTGLVFGGGTK LTVL Heavy chain SEQ ID NO: 212Light chain 270:DIELTQPPSVSVAPGQTARISCSGDSLGDYYAYWYQQKPGQAPVLVIYKDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQAWTDSPTGLVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTE X (X can be CS or A) PN encodingSEQ ID NO: 214 SEQ ID NO: 210 PN encoding271:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 269CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTCTTGGTGATTATTATGCTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGGCTTGGACTGATTCTCCTACTGGTCTTGTGTTTGGCGGCGGCACGAAGTTAACCGT TCTT PN encoding SEQ ID NO: 216 SEQID NO :212 PN encoding 272:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO:270 CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATTCTCTTGGTGATTATTATGCTTATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATAATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCCAGGCTTGGACTGATTCTCCTACTGGTCTTGTGTTTGGCGGCGGCACGAAGTTAACCGTTCTTGGCCAGCCGAAAGCCGCACCGAGTGTGACGCTGTTTCCGCCGAGCAGCGAAGAATTGCAGGCGAACAAAGCGACCCTGGTGTGCCTGATTAGCGACTTTTATCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAGGGGAGCAC CGTGGAAAAAACCGTTGCGCCGACTGAGX (Xcan be TGCAGC or GCC) Antibody 7832 CDRH1 SEQ ID NO: 17 CDRH2 SEQ ID NO:18 CDRH3 SEQ ID NO: 19 CDRL1 SEQ ID NO: 20 CDRL2 SEQ ID NO: 21 CDRL3 SEQID NO: 22 VH 273:QVQLVQSGAEVKKPGESLKISCKGSGYSFTNYISVWRQMPGKGLEWMGIIDPDDSYTEYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYEYGGFD IWGQGTLVTVSS VL SEQ ID NO: 219Heavy chain 274:QVQLVQSGAEVKKPGESLKISCKGSGYSFTNYISVWRQMPGKGLEWMGIIDPDDSYTEYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYEYGGFDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKK VEPKSX (X can be C, EF or CEF)Light chain SEQ ID NO: 221 PN encoding275:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 273GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTAATTATATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTGATGATTCTTATACTGAGTATTCTCCTTCTTTTCAGGGTCAGGTCACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATGAGTATGGTGGTTTTGATATTTGGGG CCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding SEQ ID NO: 223 SEQ ID NO :219 PN encoding276:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 274GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTAATTATATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTGATGATTCTTATACTGAGTATTCTCCTTCTTTTCAGGGTCAGGTCACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATGAGTATGGTGGTTTTGATATTTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCGTCGACCAAAGGTCCAAGCGTGTTTCCGCTGGCTCCGAGCAGCAAAAGCACCAGCGGCGGCACGGCTGCCCTGGGCTGCCTGGTTAAAGATTATTTCCCGGAACCAGTCACCGTGAGCTGGAACAGCGGGGCGCTGACCAGCGGCGTGCATACCTTTCCGGCGGTGCTGCAAAGCAGCGGCCTGTATAGCCTGAGCAGCGTTGTGACCGTGCCGAGCAGCAGCTTAGGCACTCAGACCTATATtTGCAACGTGAACCATAAACCGAGCAACACCAAAGTGGATAAAAAAGTGGA ACCGAAAAGCX (X can be TGC, GAATTCor TGCGAATTC) Antibody 7909 CDRH1 SEQ ID NO: 17 CDRH2 SEQ ID NO: 107CDRH3 SEQ ID NO: 19 CDRL1 SEQ ID NO: 20 CDRL2 SEQ ID NO: 21 CDRL3 SEQ IDNO: 22 VH 277:QVQLVQSGAEVKKPGESLKISCKGSGYSFTNYISVWRQMPGKGLEWMGIIDPQDSYTEYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYEYGGFD IWGQGTLVTVSS VL SEQ ID NO: 219Heavy chain 278:QVQLVQSGAEVKKPGESLKISCKGSGYSFTNYISVWRQMPGKGLEWMGIIDPQDSYTEYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYEYGGFDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKK VEPKSX (X can be C, EF or CEF)Light chain SEQ ID NO: 221 PN encoding279:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 277GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTAATTATATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTCAGGATTCTTATACTGAGTATTCTCCTTCTTTTCAGGGTCAGGTCACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATGAGTATGGTGGTTTTGATATTTGGGG CCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding SEQ ID NO: 223 SEQ ID NO: 219 PN encoding280:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 278GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTAATTATATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTCAGGATTCTTATACTGAGTATTCTCCTTCTTTTCAGGGTCAGGTCACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATGAGTATGGTGGTTTTGATATTTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCGTCGACCAAAGGTCCAAGCGTGTTTCCGCTGGCTCCGAGCAGCAAAAGCACCAGCGGCGGCACGGCTGCCCTGGGCTGCCTGGTTAAAGATTATTTCCCGGAACCAGTCACCGTGAGCTGGAACAGCGGGGCGCTGACCAGCGGCGTGCATACCTTTCCGGCGGTGCTGCAAAGCAGCGGCCTGTATAGCCTGAGCAGCGTTGTGACCGTGCCGAGCAGCAGCTTAGGCACTCAGACCTATATTTGCAACGTGAACCATAAACCGAGCAACACCAAAGTGGATAAAAAAGTGGA ACCGAAAAGCX (X can be TGC, GAATTCor TGCGAATTC) Antibody 7910 CDRH1 SEQ ID NO: 17 CDRH2 SEQ ID NO: 113CDRH3 SEQ ID NO: 19 CDRL1 SEQ ID NO: 20 CDRL2 SEQ ID NO: 21 CDRL3 SEQ IDNO: 22 VH 281:QVQLVQSGAEVKKPGESLKISCKGSGYSFTNYISVWRQMPGKGLEWMGIIDPEDSHTEYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYEYGGFD IWGQGTLVTVSS VL SEQ ID NO: 219Heavy chain 282:QVQLVQSGAEVKKPGESLKISCKGSGYSFTNYISVWRQMPGKGLEWMGIIDPEDSHTEYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARYEYGGFDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKK VEPKSX (X can be C, EF or CEF)Light chain SEQ ID NO: 221 PN encoding283:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID no: 281GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTAATTATATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTGAGGATTCTCATACTGAGTATTCTCCTTCTTTTCAGGGTCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATGAGTATGGTGGTTTTGATATTTGGGG CCAAGGCACCCTGGTGACGGTTAGCTCA PNencoding SEQ ID NO: 223 SEQ ID NO: 219 PN encoding284:CAGGTGCAATTGGTTCAGAGCGGCGCGGAAGT SEQ ID NO: 282GAAAAAACCGGGCGAAAGCCTGAAAATTAGCTGCAAAGGTTCCGGATATTCCTTTACTAATTATATTTCTTGGGTGCGCCAGATGCCTGGGAAGGGTCTCGAGTGGATGGGCATTATTGATCCTGAGGATTCTCATACTGAGTATTCTCCTTCTTTTCAGGGTCAGGTGACCATTAGCGCGGATAAAAGCATTAGCACCGCGTATCTTCAATGGAGCAGCCTGAAAGCGAGCGATACGGCCATGTATTATTGCGCGCGTTATGAGTATGGTGGTTTTGATATTTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCGTCGACCAAAGGTCCAAGCGTGTTTCCGCTGGCTCCGAGCAGCAAAAGCACCAGCGGCGGCACGGCTGCCCTGGGCTGCCTGGTTAAAGATTATTTCCCGGAACCAGTCACCGTGAGCTGGAACAGCGGGGCGCTGACCAGCGGCGTGCATACCTTTCCGGCGGTGCTGCAAAGCAGCGGCCTGTATAGCCTGAGCAGCGTTGTGACCGTGCCGAGCAGCAGCTTAGGCACTCAGACCTATATTTGCAACGTGAACCATAAACCGAGCAACACCAAAGTGGATAAAAAAGTGGA ACCGAAAAGCX (X can be TGC, GAATTCor TGCGAATTC) Antibody 7875 CDRH1 SEQ ID NO: 17 CDRH2 SEQ ID NO: 49CDRH3 SEQ ID NO: 19 CDRL1 SEQ ID NO: 20 CDRL2 SEQ ID NO: 21 CDRL3 SEQ IDNO: 50 VH SEQ ID NO: 218 VL 285:DIELTQPPSVSVAPGQTARISCSGDNIGNSYVHWYQQKPGQAPVLVIYKDNDRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCATWGSEDQVFGGGTKLT VL Heavy chain SEQ ID NO: 220 Lightchain 286:DIELTQPPSVSVAPGQTARISCSGDNIGNSYVHWYQQKPGQAPVLVIYKDNDRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCATWGSEDQVFGGGTKLTVLGQPKMPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSY LSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTEX(X can be CS or A) PN encoding SEQ ID NO: 222 SEQ ID NO :218 PN encoding287:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 285CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATAATATTGGTAATTCTTATGTTCATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATGATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCGCTACTTGGGGTTCTGAGGATCA GGTGTTTGGCGGCGGCACGAAGTTAACCGTTCTTPN encoding SEQ ID NO: 224 SEQ ID NO: 220 PN encoding288:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 286CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATAATATTGGTAATTCTTATGTTCATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATGATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCGCTACTTGGGGTTCTGAGGATCAGGTGTTTGGCGGCGGCACGAAGTTAACCGTTCTTGGCCAGCCGAAAGCCGCACCGAGTGTGACGCTGTTTCCGCCGAGCAGCGAAGAATTGCAGGCGAACAAAGCGACCCTGGTGTGCCTGATTAGCGACTTTTATCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAGGGGAGCACCGTGGA AAAAACCGTTGCGCCGACTGAGX (X can beTGCAGC or GCC) Antibody 7878 CDRH1 SEQ ID NO: 17 CDRH2 SEQ ID NO: 49CDRH3 SEQ ID NO: 19 CDRL1 SEQ ID NO: 20 CDRL2 SEQ ID NO: 21 CDRL3 SEQ IDNO: 101 VH SEQ ID NO: 218 VL 289:DIELTQPPSVSVAPGQTARISCSGDNIGNSYVHWYQQKPGQAPVLVIYKDNDRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCSTWDIEPTYVFGGGTKL TVL Heavy chain SEQ ID NO: 220Light chain 290:DIELTQPPSVSVAPGQTARISCSGDNIGNSYVHWYQQKPGQAPVLVIYKDNDRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCSTWDIEPTYVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTEX (X can be CS or A) PN encoding SEQID NO: 222 SEQ ID NO: 218 PN encoding291:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 289CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATAATATTGGTAATTCTTATGTTCATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATGATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCTCTACTTGGGATATTGAGCCTACTTATGTGTTTGGCGGCGGCACGAAGTTAACCGTTCT T PN encoding SEQ ID NO: 224 SEQ IDNO: 220 PN encoding 292:GATATCGAACTGACCCAGCCGCCTTCAGTGAG SEQ ID NO: 290CGTTGCACCAGGTCAGACCGCGCGTATCTCGTGTAGCGGCGATAATATTGGTAATTCTTATGTTCATTGGTACCAGCAGAAACCCGGGCAGGCGCCAGTTCTTGTGATTTATAAGGATAATGATCGTCCCTCAGGCATCCCGGAACGCTTTAGCGGATCCAACAGCGGCAACACCGCGACCCTGACCATTAGCGGCACTCAGGCGGAAGACGAAGCGGATTATTATTGCTCTACTTGGGATATTGAGCCTACTTATGTGTTTGGCGGCGGCACGAAGTTAACCGTTCTTGGCCAGCCGAAAGCCGCACCGAGTGTGACGCTGTTTCCGCCGAGCAGCGAAGAATTGCAGGCGAACAAAGCGACCCTGGTGTGCCTGATTAGCGACTTTTATCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAGGGGAGCACCGT GGAAAAAACCGTTGCGCCGACTGAGX (X canbe TGCAGC or GCC) Human 296:MGLLGILCFLIFLGKTWGQEQTYVISAPKIFR (Homosapiens) VGASENIVIQVYGYTEAFDATISIKSYPDKKFSYSS C5GHVHLSSENKFQNSAILTIQPKQLPGGQNPVSYVYLEVVSKHFSKSKRMPITYONGFLFIHTOKPVYTPOQSVKVRVYSLNOOLKPAKRETVLTFIDPEGSEVOMVEEIDHIGIISFPDFKIPSNPRYGMWTIKAKYKEDFSTTGTAYFEVKEYVLPHFSVSIEPEYNFIGYKNFKNFEITIKARYFYNKVVTEADVYITFGIREDLKDDQKEMMQTAMQNTMLINGIAQVTFDSETAVKELSYYSLEDLNNKYLYIAVTVIESTGGFSEEAEIPGIKYVLSPYKLNLVATPLFLKPGIPYPIKVQVKDSLDQLVGGVPVTLNAQTIDVNQETSDLDPSKSVTRVDDGVASFVLNLPSGVTVLEFNVKTDAPDLPEENQAREGYRAIAYSSLSQSYLYIDWTDNHKALLVGEHLNIIVTPKSPYIDKITHYNYLILSKGKIIHFGTREKFSDASYQSINIPVTQNMVPSSRLLVYYIVTGEQTAELVSDSVWLNIEEKCGNQLQVHLSPDADAYSPGQTVSLNMATGMDSWVALAAVDSAVYGVQRGAKKPLERVFQFLEKSDLGCGAGGGLNNANVFHLAGLTFLTNANADOSQENDEPCKEILRPRRTLQKKIEEIAAKYKHSVVKKCCYDGACVNNDETCEQRAARISLGPRCIKAFTECCVVASQLRANISHKDMQLGRLHMKTLLPVSKPEIRSYFPESWLWEVHLVPRRKQLQFALPDSLTTWEIQGVGISNTGICVADTVKAKVFKDVFLEMNIPYSVVRGEQIQLKGTVYNYRTSGMQFCVKMSAVEGICTSESPVIDHQGTKSSKCVRQKVEGSSSHLVTFTVLPLEIGLHNINFSLETWFGKEILVKTLRVVPEGVKRESYSGVTLDPRGIYGTISRRKEFPYRIPLDLVPKTEIKRILSVKGLLVGEILSAVLSQEGINILTHLPKGSAEAELMSVVPVFYVFHYLETGNHWNIFHSDPLIEKQKLKKKLKEGMLSIMSYRNADYSYSVWKGGSASTWLTAFALRVLGQVNKYVEQNQNSICNSLLWLVENYQLDNGSFKENSQYQPIKLQGTLPVEARENSLYLTAFTVIGIRKAFDICPLVKIDTALIKADNFLLENTLPAQSTFTLAISAYALSLGDKTHPQFRSIVSALKREALVKGNPPIYRFWKDNLQHKDSSVPNTGTARMVETTAYALLTSLNLKDINYVNPVIKWLSEEQRYGGGFYSTQDTINAIEGLTEYSLLVKQLRLSMDIDVSYKHKGALHNYKMTDKNFLGRPVEVLLNDDLIVSTGFGSGLATVHVTTVVHKTSTSEEVCSFYLKIDTQDIEASHYRGYGNSDYKRIVACASYKPSREESSSGSSHAVMDISLPTGISANEEDLKALVEGVDQLFTDYQIKDGHVILQLNSIPSSDFLCVRFRIFELFEVGFLSPATFTVYEYHRPDKQCTMFYSTSNIKIQKVCEGAACKCVEADCGQMQEELDLTISAETRKQTACKPEIAYAYKVSITSITVENVFVKYKATLLDIYKTGEAVAEKOSEITFIKKVTCTNAELVKGRQYLIMGKEALQIKYNFSFRYIYPLDSLTWIEYWPRDT TCSSCQAFLANLDEFAEDIFLNGC Cynomolgus297:MGLLGILCFLIFLGKTWGQEQTYVISAPKIFR MacaqueVGASENIVIQVYGYTEAFDATSIKSYPDKKFSYSSG (MacacaHVHLSSENKFQNSAVLTIQPKQLPGGQNQVSYVYLE fascicularis)VVSKHFSKSKKIPITYDNGFLFIHTDKPVYTPDQSV C5KVRVYSLNDDLKPAKRETVLTFIDPEGSEIDMVEEIDHIGIISFPDFKIPSNPRYGMWTIQAKYKEDFSTTGTAFFEVKEYVLPHFSVSVEPESNFIGYKNFKNFEITIKARYFYNKVVTEADVYITFGIREDLKDDQKEMMQTAMQNTMLINGIAQVTFDSETAVKELSYYSLEDLNNKYLYIAVTVIESTGGFSEEAEIPGIKYVLSPYKLNLVATPLFLKPGIPYSIKVQVKDALDQLVGGVPVTLNAQTIDVNQETSDLEPRKSVTRVDDGVASFVVNLPSGVTVLEFNVKTDAPDLPDENQAREGYRAIAYSSLSQSYLYIDWTDNHKALLVGEYLNIIVTPKSPYIDKITHYNYLILSKGKIIHFGTREKLSDASYQSINIPVTQNMVPSSRLLVYYIVTGEQTAELVSDSVWLNIEEKCGNQLQVHLSPDADTYSPGQTVSLNMVTGMDSWVALTAVDSAVYGVQRRAKKPLERVFQFLEKSDLGCGAGGGLNNANVFHLAGLTFLTNANADDSQENDEPCKEIIRPRRMLQEKIEEIAAKYKHLVVKKCCYDGVRINHDETCEQRAARISVGPRCVKAFTECCVVASQLRANNSHKDLQLGRLHMKTLLPVSKPEIRSYFPESWLWEVHLVPRRKQLQFALPDSVTTWEIQGVGISNSGICVADTIKAKVFKDVFLEMNIPYSVVRGEQVQLKGTVYNYRTSGMQFCVKMSAVEGICTSESPVIDHQGTKSSKCVRQKVEGSSNHLVTFTVLPLEIGLQNINFSLETSFGKEILVKSLRWPEGVKRESYSGITLDPRGIYGTISRRKEFPYRIPLDLVPKTEIKRILSVKGLLVGEILSAVLSREGINILTHLPKGSAEAELMSVVPVFYVFHYLETGNHWNIFHSDPLIEKRNLEKKLKEGMVSIMSYRNADYSYSVWKGGSASTWLTAFALRVLGQVHKYVEQNQNSICNSLLWLVENYQLDNGSFKENSQYQPIKLQGTLPVEARENSLYLTAFTVIGIRKAFDICPLVKINTALIKADTFLLENTLPAQSTFTLAISAYALSLGDKTHPQFRSIVSALKREALVKGNPPIYRFWKDSLQHKDSSVPNTGTARMVETTAYALLTSLNLKDINYVNPIIKWLSEEQRYGGGFYSTQDTINAIEGLTEYSLLVKQLRLNMDIDVAYKHKGPLHNYKMTDKNFLGRPVEVLLNDDLVVSTGFGSGLATVHVTVVVHKTSTSEEVCSFYLKIDTQDIEASHYRGYGNSDYKRIVACASYKPSKEESSSGSSHAVMDISLPTGINANEEDLKALVEGVDQLFTDYQIKDGHVILQLNSIPSSDFLCVRFRIFELFEVGFLSPATFTVYEYHRPDKQCTMFYSTSNIKIQKVCEGATCKCIEADCGQMQKELDLTISAETRKQTACNPEIAYAYKVIITSITVENVFVKYKATLLDIYKTGEAVAEKDSEITFIKKVTCTNAELVKGRQYLIMGKEALQIKYNFTFRYIYPLDSLTWIEYWPRDTTC SSCQAFLANLDEFAEDIFLNGC

Other antibodies of the invention include those where the amino acids ornucleic acids encoding the amino acids have been mutated, yet have atleast 60, 70, 80, 90 or 95 percent identity to the sequences describedin Table 1. In some embodiments, it include mutant amino acid sequenceswherein no more than 1, 2, 3, 4 or 5 amino acids have been mutated inthe variable regions when compared with the variable regions depicted inthe sequence described in Table 1, while retaining substantially thesame therapeutic activity.

Since each of these antibodies can bind to C5, the VH, VL, full lengthlight chain, and full length heavy chain sequences (amino acid sequencesand the nucleotide sequences encoding the amino acid sequences) can be“mixed and matched” to create other C5-binding antibodies of theinvention. Such “mixed and matched” C5-binding antibodies can be testedusing the binding assays known in the art (e.g., ELISAs, and otherassays described in the Example section). When these chains are mixedand matched, a VH sequence from a particular VH/VL pairing should bereplaced with a structurally similar VH sequence. Likewise a full lengthheavy chain sequence from a particular full length heavy chain/fulllength light chain pairing should be replaced with a structurallysimilar full length heavy chain sequence. Likewise, a VL sequence from aparticular VH/VL pairing should be replaced with a structurally similarVL sequence. Likewise a full length light chain sequence from aparticular full length heavy chain/full length light chain pairingshould be replaced with a structurally similar full length light chainsequence. Accordingly, in one aspect, the invention provides an isolatedmonoclonal antibody or antigen binding region thereof having: a heavychain variable region comprising an amino acid sequence selected fromthe group consisting of SEQ ID NOs: 7, 23, 39, 51, 67, 79, 96, 108, 114,121, 137, 151, 165, 179, 187, 201, 210, 218, 227, 241, 253, 257, 273,277, and 281; and a light chain variable region comprising an amino acidsequence selected from the group consisting of SEQ ID NOs: 8, 24, 40,52, 68, 80, 90, 102, 122, 138, 152, 166, 180, 188, 202, 211, 219, 228,242, 261, 265, 269, 285, and 289; wherein the antibody specificallybinds to C5 (e.g., human and/or cynomologus C5).

In another aspect, the invention provides (i) an isolated monoclonalantibody having: a full length heavy chain comprising an amino acidsequence that has been optimized for expression in the cell of amammalian selected from the group consisting of SEQ ID NOs: 9, 25, 41,53, 69, 81, 97, 109, 115, 123, 139, 153, 167, 181, 189, 203, 212, 220,229, 243, 249, 254, 258, 274, 278, and 282; and a full length lightchain comprising an amino acid sequence that has been optimized forexpression in the cell of a mammalian selected from the group consistingof SEQ ID NOs: 10, 26, 42, 54, 70, 82, 91, 103, 124, 140, 154, 168, 182,190, 204, 213, 221, 230, 244, 251, 262, 266, 270, 286, and 290; or (ii)a functional protein comprising an antigen binding portion thereof.

In another aspect, the present invention provides C5-binding antibodiesthat comprise the heavy chain and light chain CDR1s, CDR2s and CDR3s asdescribed in Table 1, or combinations thereof. The amino acid sequencesof the VH CDR1s of the antibodies are shown in SEQ ID NOs: 1, 17, 33,61, 131, 145, 159, 173, 195, and 235. The amino acid sequences of the VHCDR2s of the antibodies and are shown in SEQ ID NOs: 2, 18, 34, 49, 62,77, 95, 107, 113, 119, 132, 146, 160, 174, 196, 226, and 236. The aminoacid sequences of the VH CDR3s of the antibodies are shown in SEQ IDNOs: 3, 19, 35, 63, 133, 147, 161, 175, 197, and 237. The amino acidsequences of the VL CDR1s of the antibodies are shown in SEQ ID NOs: 4,20, 36, 64, 134, 148, 162, 176, 198, and 238. The amino acid sequencesof the VL CDR2s of the antibodies are shown in SEQ ID NOs: 5, 21, 37,65, 135, 149, 163, 177, 199, and 239. The amino acid sequences of the VLCDR3s of the antibodies are shown in SEQ ID NOs: 6, 22, 38, 50, 66, 78,89, 101, 120, 136, 150, 164, 178, 200, 209, and 240. The CDR regions aredelineated using the Kabat system (Kabat, E. A., et al., 1991 Sequencesof Proteins of Immunological Interest, Fifth Edition, U.S. Department ofHealth and Human Services, NIH Publication No. 91-3242).

Given that each of these antibodies can bind to C5 and thatantigen-binding specificity is provided primarily by the CDR1, 2 and 3regions, the VH CDR1, 2 and 3 sequences and VL CDR1, 2 and 3 sequencescan be “mixed and matched” (i.e., CDRs from different antibodies can bemixed and match, although each antibody must contain a VH CDR1, 2 and 3and a VL CDR1, 2 and 3 to create other C5-binding binding molecules ofthe invention. Such “mixed and matched” C5-binding antibodies can betested using the binding assays known in the art and those described inthe Examples (e.g., ELISAs). When VH CDR sequences are mixed andmatched, the CDR1, CDR2 and/or CDR3 sequence from a particular VHsequence should be replaced with a structurally similar CDR sequence(s).Likewise, when VL CDR sequences are mixed and matched, the CDR1, CDR2and/or CDR3 sequence from a particular VL sequence should be replacedwith a structurally similar CDR sequence(s). It will be readily apparentto the ordinarily skilled artisan that novel VH and VL sequences can becreated by substituting one or more VH and/or VL CDR region sequenceswith structurally similar sequences from the CDR sequences shown hereinfor monoclonal antibodies of the present invention.

Accordingly, the present invention provides an isolated monoclonalantibody or antigen binding region thereof comprising a heavy chainvariable region CDR1 comprising an amino acid sequence selected from thegroup consisting of SEQ ID NOs: 1, 17, 33, 61, 131, 145, 159, 173, 195,and 235; a heavy chain variable region CDR2 comprising an amino acidsequence selected from the group consisting of SEQ ID NOs: 2, 18, 34,49, 62, 77, 95, 107, 113, 119, 132, 146, 160, 174, 196, 226, and 236; aheavy chain variable region CDR3 comprising an amino acid sequenceselected from the group consisting of SEQ ID NOs: 3, 19, 35, 63, 133,147, 161, 175, 197, and 237; a light chain variable region CDR1comprising an amino acid sequence selected from the group consisting ofSEQ ID NOs: 4, 20, 36, 64, 134, 148, 162, 176, 198, and 238; a lightchain variable region CDR2 comprising an amino acid sequence selectedfrom the group consisting of SEQ ID NOs: 5, 21, 37, 65, 135, 149, 163,177, 199, and 239; and a light chain variable region CDR3 comprising anamino acid sequence selected from the group consisting of SEQ ID NOs: 6,22, 38, 50, 66, 78, 89, 101, 120, 136, 150, 164, 178, 200, 209, and 240;wherein the antibody specifically binds C5.

In a specific embodiment, an antibody that specifically binds to C5comprising a heavy chain variable region CDR1 of SEQ ID NO:1; a heavychain variable region CDR2 of SEQ ID NO: 2; a heavy chain variableregion CDR3 of SEQ ID NO: 3; a light chain variable region CDR1 of SEQID NO: 4; a light chain variable region CDR2 of SEQ ID NO: 5; and alight chain variable region CDR3 of SEQ ID NO: 6. In another specificembodiment, an antibody that specifically binds to C5 comprising a heavychain variable region CDR1 of SEQ ID NO: 17; a heavy chain variableregion CDR2 of SEQ ID NO: 18; a heavy chain variable region CDR3 of SEQID NO: 19; a light chain variable region CDR1 of SEQ ID NO: 20; a lightchain variable region CDR2 of SEQ ID NO: 21; and a light chain variableregion CDR3 of SEQ ID NO: 22.

In another specific embodiment, an antibody that specifically binds toC5 comprising a heavy chain variable region CDR1 of SEQ ID NO: 33; aheavy chain variable region CDR2 of SEQ ID NO: 34; a heavy chainvariable region CDR3 of SEQ ID NO: 35; a light chain variable regionCDR1 of SEQ ID NO: 36; a light chain variable region CDR2 of SEQ ID NO:37; and a light chain variable region CDR3 of SEQ ID NO: 38. In anotherspecific embodiment, an antibody that specifically binds to C5comprising a heavy chain variable region CDR1 of SEQ ID NO: 17; a heavychain variable region CDR2 of SEQ ID NO: 49; a heavy chain variableregion CDR3 of SEQ ID NO: 19; a light chain variable region CDR1 of SEQID NO: 20; a light chain variable region CDR2 of SEQ ID NO: 21; and alight chain variable region CDR3 of SEQ ID NO: 50.

In another specific embodiment, an antibody that specifically binds toC5 comprising a heavy chain variable region CDR1 of SEQ ID NO: 61; aheavy chain variable region CDR2 of SEQ ID NO: 62; a heavy chainvariable region CDR3 of SEQ ID NO: 63; a light chain variable regionCDR1 of SEQ ID NO: 64; a light chain variable region CDR2 of SEQ ID NO:65; and a light chain variable region CDR3 of SEQ ID NO: 66. In anotherspecific embodiment, an antibody that specifically binds to C5comprising a heavy chain variable region CDR1 of SEQ ID NO: 61; a heavychain variable region CDR2 of SEQ ID NO: 77; a heavy chain variableregion CDR3 of SEQ ID NO: 63; a light chain variable region CDR1 of SEQID NO: 64; a light chain variable region CDR2 of SEQ ID NO: 65; and alight chain variable region CDR3 of SEQ ID NO: 78.

In another specific embodiment, an antibody that specifically binds toC5 comprising a heavy chain variable region CDR1 of SEQ ID NO: 61; aheavy chain variable region CDR2 of SEQ ID NO: 77; a heavy chainvariable region CDR3 of SEQ ID NO: 63; a light chain variable regionCDR1 of SEQ ID NO: 64; a light chain variable region CDR2 of SEQ ID NO:65; and a light chain variable region CDR3 of SEQ ID NO: 89. In anotherspecific embodiment, an antibody that specifically binds to C5comprising a heavy chain variable region CDR1 of SEQ ID NO: 61; a heavychain variable region CDR2 of SEQ ID NO: 62; a heavy chain variableregion CDR3 of SEQ ID NO: 63; a light chain variable region CDR1 of SEQID NO: 64; a light chain variable region CDR2 of SEQ ID NO: 65; and alight chain variable region CDR3 of SEQ ID NO: 89.

In another specific embodiment, an antibody that specifically binds toC5 comprising a heavy chain variable region CDR1 of SEQ ID NO: 61; aheavy chain variable region CDR2 of SEQ ID NO: 95; a heavy chainvariable region CDR3 of SEQ ID NO: 63; a light chain variable regionCDR1 of SEQ ID NO: 64; a light chain variable region CDR2 of SEQ ID NO:65; and a light chain variable region CDR3 of SEQ ID NO: 89. In anotherspecific embodiment, an antibody that specifically binds to C5comprising a heavy chain variable region CDR1 of SEQ ID NO: 17; a heavychain variable region CDR2 of SEQ ID NO: 49; a heavy chain variableregion CDR3 of SEQ ID NO: 19; a light chain variable region CDR1 of SEQID NO: 20; a light chain variable region CDR2 of SEQ ID NO: 21; and alight chain variable region CDR3 of SEQ ID NO: 101.

In another specific embodiment, an antibody that specifically binds toC5 comprising a heavy chain variable region CDR1 of SEQ ID NO: 17; aheavy chain variable region CDR2 of SEQ ID NO: 107; a heavy chainvariable region CDR3 of SEQ ID NO: 19; a light chain variable regionCDR1 of SEQ ID NO: 20; a light chain variable region CDR2 of SEQ ID NO:21; and a light chain variable region CDR3 of SEQ ID NO: 22. In anotherspecific embodiment, an antibody that specifically binds to C5comprising a heavy chain variable region CDR1 of SEQ ID NO: 17; a heavychain variable region CDR2 of SEQ ID NO: 107; a heavy chain variableregion CDR3 of SEQ ID NO: 19; a light chain variable region CDR1 of SEQID NO: 20; a light chain variable region CDR2 of SEQ ID NO: 21; and alight chain variable region CDR3 of SEQ ID NO: 101.

In another specific embodiment, an antibody that specifically binds toC5 comprising a heavy chain variable region CDR1 of SEQ ID NO: 17; aheavy chain variable region CDR2 of SEQ ID NO: 113; a heavy chainvariable region CDR3 of SEQ ID NO: 19; a light chain variable regionCDR1 of SEQ ID NO: 20; a light chain variable region CDR2 of SEQ ID NO:21; and a light chain variable region CDR3 of SEQ ID NO: 22. In anotherspecific embodiment, an antibody that specifically binds to C5comprising a heavy chain variable region CDR1 of SEQ ID NO: 17; a heavychain variable region CDR2 of SEQ ID NO: 113; a heavy chain variableregion CDR3 of SEQ ID NO: 19; a light chain variable region CDR1 of SEQID NO: 20; a light chain variable region CDR2 of SEQ ID NO: 21; and alight chain variable region CDR3 of SEQ ID NO: 101.

In another specific embodiment, an antibody that specifically binds toC5 comprising a heavy chain variable region CDR1 of SEQ ID NO: 1; aheavy chain variable region CDR2 of SEQ ID NO: 119; a heavy chainvariable region CDR3 of SEQ ID NO: 3; a light chain variable region CDR1of SEQ ID NO: 4; a light chain variable region CDR2 of SEQ ID NO: 5; anda light chain variable region CDR3 of SEQ ID NO: 120. In anotherspecific embodiment, an antibody that specifically binds to C5comprising a heavy chain variable region CDR1 of SEQ ID NO: 131; a heavychain variable region CDR2 of SEQ ID NO: 132; a heavy chain variableregion CDR3 of SEQ ID NO: 133; a light chain variable region CDR1 of SEQID NO: 134; a light chain variable region CDR2 of SEQ ID NO: 135; and alight chain variable region CDR3 of SEQ ID NO: 136.

In another specific embodiment, an antibody that specifically binds toC5 comprising a heavy chain variable region CDR1 of SEQ ID NO: 145; aheavy chain variable region CDR2 of SEQ ID NO: 146; a heavy chainvariable region CDR3 of SEQ ID NO: 147; a light chain variable regionCDR1 of SEQ ID NO: 148; a light chain variable region CDR2 of SEQ ID NO:149; and a light chain variable region CDR3 of SEQ ID NO: 150. Inanother specific embodiment, an antibody that specifically binds to C5comprising a heavy chain variable region CDR1 of SEQ ID NO: 159; a heavychain variable region CDR2 of SEQ ID NO: 160; a heavy chain variableregion CDR3 of SEQ ID NO: 161; a light chain variable region CDR1 of SEQID NO: 162; a light chain variable region CDR2 of SEQ ID NO: 163; and alight chain variable region CDR3 of SEQ ID NO: 164.

In another specific embodiment, an antibody that specifically binds toC5 comprising a heavy chain variable region CDR1 of SEQ ID NO: 173; aheavy chain variable region CDR2 of SEQ ID NO: 174; a heavy chainvariable region CDR3 of SEQ ID NO: 175; a light chain variable regionCDR1 of SEQ ID NO: 176; a light chain variable region CDR2 of SEQ ID NO:177; and a light chain variable region CDR3 of SEQ ID NO: 178. Inanother specific embodiment, an antibody that specifically binds to C5comprising a heavy chain variable region CDR1 of SEQ ID NO: 195; a heavychain variable region CDR2 of SEQ ID NO: 196; a heavy chain variableregion CDR3 of SEQ ID NO: 197; a light chain variable region CDR1 of SEQID NO: 198; a light chain variable region CDR2 of SEQ ID NO: 199; and alight chain variable region CDR3 of SEQ ID NO: 200.

In another specific embodiment, an antibody that specifically binds toC5 comprising a heavy chain variable region CDR1 of SEQ ID NO: 61; aheavy chain variable region CDR2 of SEQ ID NO: 77; a heavy chainvariable region CDR3 of SEQ ID NO: 63; a light chain variable regionCDR1 of SEQ ID NO: 64; a light chain variable region CDR2 of SEQ ID NO:65; and a light chain variable region CDR3 of SEQ ID NO: 209. In anotherspecific embodiment, an antibody that specifically binds to C5comprising a heavy chain variable region CDR1 of SEQ ID NO: 17; a heavychain variable region CDR2 of SEQ ID NO: 49; a heavy chain variableregion CDR3 of SEQ ID NO: 19; a light chain variable region CDR1 of SEQID NO: 20; a light chain variable region CDR2 of SEQ ID NO: 21; and alight chain variable region CDR3 of SEQ ID NO: 22.

In another specific embodiment, an antibody that specifically binds toC5 comprising a heavy chain variable region CDR1 of SEQ ID NO: 33; aheavy chain variable region CDR2 of SEQ ID NO: 226; a heavy chainvariable region CDR3 of SEQ ID NO: 35; a light chain variable regionCDR1 of SEQ ID NO: 36; a light chain variable region CDR2 of SEQ ID NO:37; and a light chain variable region CDR3 of SEQ ID NO: 38. In anotherspecific embodiment, an antibody that specifically binds to C5comprising a heavy chain variable region CDR1 of SEQ ID NO: 235; a heavychain variable region CDR2 of SEQ ID NO: 236; a heavy chain variableregion CDR3 of SEQ ID NO: 237; a light chain variable region CDR1 of SEQID NO: 238; a light chain variable region CDR2 of SEQ ID NO: 239; and alight chain variable region CDR3 of SEQ ID NO: 240.

In another specific embodiment, an antibody that specifically binds toC5 comprising a heavy chain variable region CDR1 of SEQ ID NO: 1; aheavy chain variable region CDR2 of SEQ ID NO: 119; a heavy chainvariable region CDR3 of SEQ ID NO: 3; a light chain variable region CDR1of SEQ ID NO: 4; a light chain variable region CDR2 of SEQ ID NO: 5; anda light chain variable region CDR3 of SEQ ID NO: 6. In another specificembodiment, an antibody that specifically binds to C5 comprising a heavychain variable region CDR1 of SEQ ID NO: 1; a heavy chain variableregion CDR2 of SEQ ID NO: 2; a heavy chain variable region CDR3 of SEQID NO: 3; a light chain variable region CDR1 of SEQ ID NO: 4; a lightchain variable region CDR2 of SEQ ID NO: 5; and a light chain variableregion CDR3 of SEQ ID NO: 120.

In another specific embodiment, an antibody that specifically binds toC5 comprising a heavy chain variable region CDR1 of SEQ ID NO: 61; aheavy chain variable region CDR2 of SEQ ID NO: 62; a heavy chainvariable region CDR3 of SEQ ID NO: 63; a light chain variable regionCDR1 of SEQ ID NO: 64; a light chain variable region CDR2 of SEQ ID NO:65; and a light chain variable region CDR3 of SEQ ID NO: 209. In anotherspecific embodiment, an antibody that specifically binds to C5comprising a heavy chain variable region CDR1 of SEQ ID NO: 61; a heavychain variable region CDR2 of SEQ ID NO: 95; a heavy chain variableregion CDR3 of SEQ ID NO: 63; a light chain variable region CDR1 of SEQID NO: 64; a light chain variable region CDR2 of SEQ ID NO: 65; and alight chain variable region CDR3 of SEQ ID NO: 209.

In another specific embodiment, an antibody that specifically binds toC5 comprising a heavy chain variable region CDR1 of SEQ ID NO: 61; aheavy chain variable region CDR2 of SEQ ID NO: 77; a heavy chainvariable region CDR3 of SEQ ID NO: 63; a light chain variable regionCDR1 of SEQ ID NO: 64; a light chain variable region CDR2 of SEQ ID NO:65; and a light chain variable region CDR3 of SEQ ID NO: 66. In anotherspecific embodiment, an antibody that specifically binds to C5comprising a heavy chain variable region CDR1 of SEQ ID NO: 61; a heavychain variable region CDR2 of SEQ ID NO: 77; a heavy chain variableregion CDR3 of SEQ ID NO: 63; a light chain variable region CDR1 of SEQID NO: 64; a light chain variable region CDR2 of SEQ ID NO: 65; and alight chain variable region CDR3 of SEQ ID NO: 78.

In another specific embodiment, an antibody that specifically binds toC5 comprising a heavy chain variable region CDR1 of SEQ ID NO: 61; aheavy chain variable region CDR2 of SEQ ID NO: 77; a heavy chainvariable region CDR3 of SEQ ID NO: 63; a light chain variable regionCDR1 of SEQ ID NO: 64; a light chain variable region CDR2 of SEQ ID NO:65; and a light chain variable region CDR3 of SEQ ID NO: 89. In anotherspecific embodiment, an antibody that specifically binds to C5comprising a heavy chain variable region CDR1 of SEQ ID NO: 17; a heavychain variable region CDR2 of SEQ ID NO: 107; a heavy chain variableregion CDR3 of SEQ ID NO: 19; a light chain variable region CDR1 of SEQID NO: 20; a light chain variable region CDR2 of SEQ ID NO: 21; and alight chain variable region CDR3 of SEQ ID NO: 22.

In certain embodiments, an antibody that specifically binds to C5 is anantibody that is described in Table 1.

As used herein, a human antibody comprises heavy or light chain variableregions or full length heavy or light chains that are “the product of”or “derived from” a particular germline sequence if the variable regionsor full length chains of the antibody are obtained from a system thatuses human germline immunoglobulin genes. Such systems includeimmunizing a transgenic mouse carrying human immunoglobulin genes withthe antigen of interest or screening a human immunoglobulin gene librarydisplayed on phage with the antigen of interest. A human antibody thatis “the product of” or “derived from” a human germline immunoglobulinsequence can be identified as such by comparing the amino acid sequenceof the human antibody to the amino acid sequences of human germlineimmunoglobulins and selecting the human germline immunoglobulin sequencethat is closest in sequence (i.e., greatest % identity) to the sequenceof the human antibody. A human antibody that is “the product of” or“derived from” a particular human germline immunoglobulin sequence maycontain amino acid differences as compared to the germline sequence, dueto, for example, naturally occurring somatic mutations or intentionalintroduction of site-directed mutations. However, in the VH or VLframework regions, a selected human antibody typically is at least 90%identical in amino acids sequence to an amino acid sequence encoded by ahuman germline immunoglobulin gene and contains amino acid residues thatidentify the human antibody as being human when compared to the germlineimmunoglobulin amino acid sequences of other species (e.g., murinegermline sequences). In certain cases, a human antibody may be at least60%, 70%, 80%, 90%, or at least 95%, or even at least 96%, 97%, 98%, or99% identical in amino acid sequence to the amino acid sequence encodedby the germline immunoglobulin gene. Typically, a recombinant humanantibody will display no more than 10 amino acid differences from theamino acid sequence encoded by the human germline immunoglobulin gene inthe VH or VL framework regions. In certain cases, the human antibody maydisplay no more than 5, or even no more than 4, 3, 2, or 1 amino aciddifference from the amino acid sequence encoded by the germlineimmunoglobulin gene.

Homologous Antibodies

In yet another embodiment, the present invention provides an antibody oran antigen-binding fragment thereof comprising amino acid sequences thatare homologous to the sequences described in Table 1, and said antibodybinds to a C5 protein (e.g., human and/or cynomologus C5), and retainsthe desired functional properties of those antibodies described in Table1.

For example, the invention provides an isolated monoclonal antibody (ora functional antigen binding fragment thereof) comprising a heavy chainvariable region and a light chain variable region, wherein the heavychain variable region comprises an amino acid sequence that is at least80%, at least 90%, or at lest 95% identical to an amino acid sequenceselected from the group consisting of SEQ ID NOs: 7, 23, 39, 51, 67, 79,96, 108, 114, 121, 137, 151, 165, 179, 187, 201, 210, 218, 227, 241,253, 257, 273, 277, or 281; the light chain variable region comprises anamino acid sequence that is at least 80%, at least 90%, or at least 95%identical to an amino acid sequence selected from the group consistingof SEQ ID NOs: 8, 24, 40, 52, 68, 80, 90, 102, 122, 138, 152, 166, 180,188, 202, 211, 219, 228, 242, 261, 265, 269, 285, or 289; the antibodyspecifically binds to C5 (e.g., human and/or cynomologus C5), and theantibody can inhibit red blood cell lysis in a hemolytic assay. In aspecific example, such antibodies have an IC₅₀ value in a hemolyticassay of 20-200 pM when using human C5-depleted serum that isreconstituted with 100 pM human C5.

In other embodiments, the VH and/or VL amino acid sequences may be 50%,60%, 70%, 80%, 90%, 95%, 96%, 97%, 98% or 99% identical to the sequencesset forth in Table 1. In other embodiments, the VH and/or VL amino acidsequences may be identical except an amino acid substitution in no morethan 1, 2, 3, 4 or 5 amino acid position. An antibody having VH and VLregions having high (i.e., 80% or greater) identity to the VH and VLregions of those described in Table 1 can be obtained by mutagenesis(e.g., site-directed or PCR-mediated mutagenesis) of nucleic acidmolecules encoding SEQ ID NOs: 7, 23, 39, 51, 67, 79, 96, 108, 114, 121,137, 151, 165, 179, 187, 201, 210, 218, 227, 241, 253, 257, 273, 277, or281; and 8, 24, 40, 52, 68, 80, 90, 102, 122, 138, 152, 166, 180, 188,202, 211, 219, 228, 242, 261, 265, 269, 285, or 289 respectively,followed by testing of the encoded altered antibody for retainedfunction using the functional assays described herein.

In other embodiments, the full length heavy chain and/or full lengthlight chain amino acid sequences may be 50% 60%, 70%, 80%, 90%, 95%,96%, 97%, 98% or 99% identical to the sequences set forth in Table 1. Anantibody having a full length heavy chain and full length light chainhaving high (i.e., 80% or greater) identity to the full length heavychains of any of SEQ ID NOs: 9, 25, 41, 53, 69, 81, 97, 109, 115, 123,139, 153, 167, 181, 189, 203, 212, 220, 229, 243, 249, 254, 258, 274,278, and 282 and full length light chains of any of SEQ ID NOs 10, 26,42, 54, 70, 82, 91, 103, 124, 140, 154, 168, 182, 190, 204, 213, 221,230, 244, 251, 262, 266, 270, 286, and 290 respectively, can be obtainedby mutagenesis (e.g., site-directed or PCR-mediated mutagenesis) ofnucleic acid molecules encoding such polypeptides respectively, followedby testing of the encoded altered antibody for retained function usingthe functional assays described herein.

In other embodiments, the full length heavy chain and/or full lengthlight chain nucleotide sequences may be 60%, 70%, 80%, 90%, 95%, 96%,97%, 98% or 99% identical to the sequences set forth above.

In other embodiments, the variable regions of heavy chain and/or lightchain nucleotide sequences may be 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%or 99% identical to the sequences set forth above

As used herein, the percent identity between the two sequences is afunction of the number of identical positions shared by the sequences(i.e., % identity equals number of identical positions/total number ofpositions×100), taking into account the number of gaps, and the lengthof each gap, which need to be introduced for optimal alignment of thetwo sequences. The comparison of sequences and determination of percentidentity between two sequences can be accomplished using a mathematicalalgorithm, as described in the non-limiting examples below.

Additionally or alternatively, the protein sequences of the presentinvention can further be used as a “query sequence” to perform a searchagainst public databases to, for example, identify related sequences.For example, such searches can be performed using the BLAST program(version 2.0) of Altschul, et al., 1990 J. Mol. Biol. 215:403-10.

Antibodies with Conservative Modifications

In certain embodiments, an antibody of the invention has a heavy chainvariable region comprising CDR1, CDR2, and CDR3 sequences and a lightchain variable region comprising CDR1, CDR2, and CDR3 sequences, whereinone or more of these CDR sequences have specified amino acid sequencesbased on the antibodies described herein or conservative modificationsthereof, and wherein the antibodies retain the desired functionalproperties of the C5-binding antibodies of the invention. Accordingly,the invention provides an isolated monoclonal antibody, or a functionalantigen binding fragment thereof, consisting of a heavy chain variableregion comprising CDR1, CDR2, and CDR3 sequences and a light chainvariable region comprising CDR1, CDR2, and CDR3 sequences, wherein: theheavy chain variable region CDR1 amino acid sequences are selected fromthe group consisting of SEQ ID NOs: 1, 17, 33, 61, 131, 145, 159, 173,195, and 235, and conservative modifications thereof; the heavy chainvariable region CDR2 amino acid sequences are selected from the groupconsisting of SEQ ID NOs: 2, 18, 34, 49, 62, 77, 95, 107, 113, 119, 132,146, 160, 174, 196, 226, and 236, and conservative modificationsthereof; the heavy chain variable region CDR3 amino acid sequences areselected from the group consisting of SEQ ID NOs: 3, 19, 35, 63, 133,147, 161, 175, 197, and 237, and conservative modifications thereof; thelight chain variable regions CDR1 amino acid sequences are selected fromthe group consisting of SEQ ID NOs: 4, 20, 36, 64, 134, 148, 162, 176,198, and 238, and conservative modifications thereof; the light chainvariable regions CDR2 amino acid sequences are selected from the groupconsisting of SEQ ID NOs: 5, 21, 37, 65, 135, 149, 163, 177, 199, and239, and conservative modifications thereof; the light chain variableregions of CDR3 amino acid sequences are selected from the groupconsisting of SEQ ID NOs: 6, 22, 38, 50, 66, 78, 89, 101, 120, 136, 150,164, 178, 200, 209, and 240, and conservative modifications thereof; theantibody or the antigen-binding fragment thereof specifically binds toC5, and inhibits red blood cell lysis in a hemolytic assay as describedherein.

In other embodiments, an antibody of the invention optimized forexpression in a mammalian cell has a full length heavy chain sequenceand a full length light chain sequence, wherein one or more of thesesequences have specified amino acid sequences based on the antibodiesdescribed herein or conservative modifications thereof, and wherein theantibodies retain the desired functional properties of the C5-bindingantibodies of the invention. Accordingly, the invention provides anisolated monoclonal antibody optimized for expression in a mammaliancell consisting of a full length heavy chain and a full length lightchain wherein: the full length heavy chain has amino acid sequencesselected from the group of SEQ ID NOs: 9, 25, 41, 53, 69, 81, 97, 109,115, 123, 139, 153, 167, 181, 189, 203, 212, 220, 229, 243, 249, 254,258, 274, 278, and 282, and conservative modifications thereof; and thefull length light chain has amino acid sequences selected from the groupof SEQ ID NOs: 10, 26, 42, 54, 70, 82, 91, 103, 124, 140, 154, 168, 182,190, 204, 213, 221, 230, 244, 251, 262, 266, 270, 286, and 290, andconservative modifications thereof; the antibody specifically binds toC5 (e.g., human and/or cynomologus C5); and the antibody inhibits redblood cell lysis in a hemolytic assay as described herein. In a specificembodiment, such antibodies have an IC₅₀ value in a hemolytic assay of20-200 pM when using human C5-depleted serum that is reconstituted with100 pM human C5.

Antibodies that Bind to the Same Epitope

The present invention provides antibodies that bind to the same epitopeas do the C5-binding antibodies described in Table 1. Additionalantibodies can therefore be identified based on their ability tocross-compete (e.g., to competitively inhibit the binding of, in astatistically significant manner) with other antibodies of the inventionin C5 binding assays. The ability of a test antibody to inhibit thebinding of antibodies of the present invention to a C5 protein (e.g.,human and/or cynomolgus C5) demonstrates that the test antibody cancompete with that antibody for binding to C5; such an antibody may,according to non-limiting theory, bind to the same or a related (e.g., astructurally similar or spatially proximal) epitope on the C5 protein asthe antibody with which it competes. In a certain embodiment, theantibody that binds to the same epitope on C5 as the antibodies of thepresent invention is a human monoclonal antibody. Such human monoclonalantibodies can be prepared and isolated as described herein.

Engineered and Modified Antibodies

An antibody of the invention further can be prepared using an antibodyhaving one or more of the VH and/or VL sequences shown herein asstarting material to engineer a modified antibody, which modifiedantibody may have altered properties from the starting antibody. Anantibody can be engineered by modifying one or more residues within oneor both variable regions (i.e., VH and/or VL), for example within one ormore CDR regions and/or within one or more framework regions.Additionally or alternatively, an antibody can be engineered bymodifying residues within the constant region(s), for example to alterthe effector function(s) of the antibody.

One type of variable region engineering that can be performed is CDRgrafting. Antibodies interact with target antigens predominantly throughamino acid residues that are located in the six heavy and light chaincomplementarity determining regions (CDRs). For this reason, the aminoacid sequences within CDRs are more diverse between individualantibodies than sequences outside of CDRs. Because CDR sequences areresponsible for most antibody-antigen interactions, it is possible toexpress recombinant antibodies that mimic the properties of specificnaturally occurring antibodies by constructing expression vectors thatinclude CDR sequences from the specific naturally occurring antibodygrafted onto framework sequences from a different antibody withdifferent properties (see, e.g., Riechmann, L. et al., 1998 Nature332:323-327; Jones, P. et al., 1986 Nature 321:522-525; Queen, C. etal., 1989 Proc. Natl. Acad., U.S.A. 86:10029-10033; U.S. Pat. No.5,225,539 to Winter, and U.S. Pat. Nos. 5,530,101; 5,585,089; 5,693,762and 6,180,370 to Queen et al.)

Accordingly, another embodiment of the invention pertains to an isolatedmonoclonal antibody, or an antigen binding fragment thereof, comprisinga heavy chain variable region comprising CDR1 sequences having an aminoacid sequence selected from the group consisting of SEQ ID NOs: 1, 17,33, 61, 131, 145, 159, 173, 195, and 235; CDR2 sequences having an aminoacid sequence selected from the group consisting of SEQ ID NOs: 2, 18,34, 49, 62, 77, 95, 107, 113, 119, 132, 146, 160, 174, 196, 226, and236; CDR3 sequences having an amino acid sequence selected from thegroup consisting of SEQ ID NOs: 3, 19, 35, 63, 133, 147, 161, 175, 197,and 237, respectively; and a light chain variable region having CDR1sequences having an amino acid sequence selected from the groupconsisting of SEQ ID NOs: 4, 20, 36, 64, 134, 148, 162, 176, 198, and238; CDR2 sequences having an amino acid sequence selected from thegroup consisting of SEQ ID NOs: 5, 21, 37, 65, 135, 149, 163, 177, 199,and 239; and CDR3 sequences consisting of an amino acid sequenceselected from the group consisting of SEQ ID NOs: 6, 22, 38, 50, 66, 78,89, 101, 120, 136, 150, 164, 178, 200, 209, and 240, respectively. Thus,such antibodies contain the VH and VL CDR sequences of monoclonalantibodies, yet may contain different framework sequences from theseantibodies.

Such framework sequences can be obtained from public DNA databases orpublished references that include germine antibody gene sequences. Forexample, germine DNA sequences for human heavy and light chain variableregion genes can be found in the “VBase” human germline sequencedatabase (available on the Internet at www.mrc-cpe.cam.ac.uk/vbase), aswell as in Kabat, E. A., et al., 1991 Sequences of Proteins ofImmunological Interest, Fifth Edition, U.S. Department of Health andHuman Services, NIH Publication No. 91-3242; Tomlinson, I. M., et al.,1992 J. fol. Biol. 227:776-798; and Cox, J. P. L. et al., 1994 Eur. JImmunol. 24:827-836; the contents of each of which are expresslyincorporated herein by reference.

An example of framework sequences for use in the antibodies of theinvention are those that are structurally similar to the frameworksequences used by selected antibodies of the invention, e.g., consensussequences and/or framework sequences used by monoclonal antibodies ofthe invention. The VH CDR1, 2 and 3 sequences, and the VL CDR1, 2 and 3sequences, can be grafted onto framework regions that have the identicalsequence as that found in the germline immunoglobulin gene from whichthe framework sequence derive, or the CDR sequences can be grafted ontoframework regions that contain one or more mutations as compared to thegermline sequences. For example, it has been found that in certaininstances it is beneficial to mutate residues within the frameworkregions to maintain or enhance the antigen binding ability of theantibody (see e.g., U.S. Pat. Nos. 5,530,101; 5,585,089; 5,693,762 and6,180,370 to Queen et al).

Another type of variable region modification is to mutate amino acidresidues within the VH and/or VL CDR1, CDR2 and/or CDR3 regions tothereby improve one or more binding properties (e.g., affinity) of theantibody of interest, known as “affinity maturation.” Site-directedmutagenesis or PCR-mediated mutagenesis can be performed to introducethe mutation(s) and the effect on antibody binding, or other functionalproperty of interest, can be evaluated in in vitro or in vivo assays asdescribed herein and provided in the Examples. Conservativemodifications (as discussed above) can be introduced. The mutations maybe amino acid substitutions, additions or deletions. Moreover, typicallyno more than one, two, three, four or five residues within a CDR regionare altered.

Accordingly, in another embodiment, the invention provides isolatedC5-binding monoclonal antibodies, or an antigen binding fragmentthereof, consisting of a heavy chain variable region having: a VH CDR1region consisting of an amino acid sequence selected from the grouphaving SEQ ID NOs: 1, 17, 33, 61, 131, 145, 159, 173, 195, and 235 or anamino acid sequence having one, two, three, four or five amino acidsubstitutions, deletions or additions as compared to SEQ ID NOs: 1, 17,33, 61, 131, 145, 159, 173, 195, and 235; a VH CDR2 region having anamino acid sequence selected from the group consisting of SEQ ID NOs: 2,18, 34, 49, 62, 77, 95, 107, 113, 119, 132, 146, 160, 174, 196, 226, and236, or an amino acid sequence having one, two, three, four or fiveamino acid substitutions, deletions or additions as compared to SEQ IDNOs: 2, 18, 34, 49, 62, 77, 95, 107, 113, 119, 132, 146, 160, 174, 196,226, and 236; a VH CDR3 region having an amino acid sequence selectedfrom the group consisting of SEQ ID NOs: 3, 19, 35, 63, 133, 147, 161,175, 197, and 237, or an amino acid sequence having one, two, three,four or five amino acid substitutions, deletions or additions ascompared to SEQ ID NOs: 3, 19, 35, 63, 133, 147, 161, 175, 197, and 237;a VL CDR1 region having an amino acid sequence selected from the groupconsisting of SEQ ID NOs: 4, 20, 36, 64, 134, 148, 162, 176, 198, and238, or an amino acid sequence having one, two, three, four or fiveamino acid substitutions, deletions or additions as compared to SEQ IDNOs: 4, 20, 36, 64, 134, 148, 162, 176, 198, and 238; a VL CDR2 regionhaving an amino acid sequence selected from the group consisting of SEQID NOs: 5, 21, 37, 65, 135, 149, 163, 177, 199, and 239, or an aminoacid sequence having one, two, three, four or five amino acidsubstitutions, deletions or additions as compared to SEQ ID NOs: 5, 21,37, 65, 135, 149, 163, 177, 199, and 239; and a VL CDR3 region having anamino acid sequence selected from the group consisting of SEQ ID NOs: 6,22, 38, 50, 66, 78, 89, 101, 120, 136, 150, 164, 178, 200, 209, and 240,or an amino acid sequence having one, two, three, four or five aminoacid substitutions, deletions or additions as compared to SEQ ID NOs: 6,22, 38, 50, 66, 78, 89, 101, 120, 136, 150, 164, 178, 200, 209, and 240.

Grafting Antigen-Binding Domains into Alternative Frameworks orScaffolds

A wide variety of antibody/immunoglobulin frameworks or scaffolds can beemployed so long as the resulting polypeptide includes at least onebinding region which specifically binds to C5. Such frameworks orscaffolds include the 5 main idiotypes of human immunoglobulins, orfragments thereof, and include immunoglobulins of other animal species,preferably having humanized aspects. Single heavy-chain antibodies suchas those identified in camelids are of particular interest in thisregard. Novel frameworks, scaffolds and fragments continue to bediscovered and developed by those skilled in the art.

In one aspect, the invention pertains to generating non-immunoglobulinbased antibodies using non-immunoglobulin scaffolds onto which CDRs ofthe invention can be grafted. Known or future non-immunoglobulinframeworks and scaffolds may be employed, as long as they comprise abinding region specific for the target C5 protein (e.g., human and/orcynomolgus C5). Known non-immunoglobulin frameworks or scaffoldsinclude, but are not limited to, fibronectin (Compound Therapeutics,Inc., Waltham, Mass.), ankyrin (Molecular Partners AG, Zurich,Switzerland), domain antibodies (Domantis, Ltd., Cambridge, Mass., andAblynx nv, Zwijnaarde, Belgium), lipocalin (Pieris Proteolab AG,Freising, Germany), small modular immuno-pharmaceuticals (TrubionPharmaceuticals Inc., Seattle, Wash.), maxybodies (Avidia, Inc.,Mountain View, Calif.), Protein A (Affibody AG, Sweden), and affilin(gamma-crystallin or ubiquitin) (SciI Proteins GmbH, Halle, Germany).

The fibronectin scaffolds are based on fibronectin type III domain(e.g., the tenth module of the fibronectin type III (10 Fn3 domain)).The fibronectin type III domain has 7 or 8 beta strands which aredistributed between two beta sheets, which themselves pack against eachother to form the core of the protein, and further containing loops(analogous to CDRs) which connect the beta strands to each other and aresolvent exposed. There are at least three such loops at each edge of thebeta sheet sandwich, where the edge is the boundary of the proteinperpendicular to the direction of the beta strands (see U.S. Pat. No.6,818,418). These fibronectin-based scaffolds are not an immunoglobulin,although the overall fold is closely related to that of the smallestfunctional antibody fragment, the variable region of the heavy chain,which comprises the entire antigen recognition unit in camel and llamaIgG. Because of this structure, the non-immunoglobulin antibody mimicsantigen binding properties that are similar in nature and affinity tothose of antibodies. These scaffolds can be used in a loop randomizationand shuffling strategy in vitro that is similar to the process ofaffinity maturation of antibodies in vivo. These fibronectin-basedmolecules can be used as scaffolds where the loop regions of themolecule can be replaced with CDRs of the invention using standardcloning techniques.

The ankyrin technology is based on using proteins with ankyrin derivedrepeat modules as scaffolds for bearing variable regions which can beused for binding to different targets. The ankyrin repeat module is a 33amino acid polypeptide consisting of two anti-parallel α-helices and aβ-turn. Binding of the variable regions is mostly optimized by usingribosome display.

Avimers are derived from natural A-domain containing protein such asLRP-1. These domains are used by nature for protein-protein interactionsand in human over 250 proteins are structurally based on A-domains.Avimers consist of a number of different “A-domain” monomers (2-10)linked via amino acid linkers. Avimers can be created that can bind tothe target antigen using the methodology described in, for example, U.S.Patent Application Publication Nos. 20040175756; 20050053973;20050048512; and 20060008844.

Affibody affinity ligands are small, simple proteins composed of athree-helix bundle based on the scaffold of one of the IgG-bindingdomains of Protein A. Protein A is a surface protein from the bacteriumStaphylococcus aureus. This scaffold domain consists of 58 amino acids,13 of which are randomized to generate affibody libraries with a largenumber of ligand variants (See e.g., U.S. Pat. No. 5,831,012). Affibodymolecules mimic antibodies, they have a molecular weight of 6 kDa,compared to the molecular weight of antibodies, which is 150 kDa. Inspite of its small size, the binding site of affibody molecules issimilar to that of an antibody.

Anticalins are products developed by the company Pieris ProteoLab AG.They are derived from lipocalins, a widespread group of small and robustproteins that are usually involved in the physiological transport orstorage of chemically sensitive or insoluble compounds. Several naturallipocalins occur in human tissues or body liquids. The proteinarchitecture is reminiscent of immunoglobulins, with hypervariable loopson top of a rigid framework. However, in contrast with antibodies ortheir recombinant fragments, lipocalins are composed of a singlepolypeptide chain with 160 to 180 amino acid residues, being justmarginally bigger than a single immunoglobulin domain. The set of fourloops, which makes up the binding pocket, shows pronounced structuralplasticity and tolerates a variety of side chains. The binding site canthus be reshaped in a proprietary process in order to recognizeprescribed target molecules of different shape with high affinity andspecificity. One protein of lipocalin family, the bilin-binding protein(BBP) of Pieris Brassicae has been used to develop anticalins bymutagenizing the set of four loops. One example of a patent applicationdescribing anticalins is in PCT Publication No. WO 199916873.

Affilin molecules are small non-immunoglobulin proteins which aredesigned for specific affinities towards proteins and small molecules.New affilin molecules can be very quickly selected from two libraries,each of which is based on a different human derived scaffold protein.Affilin molecules do not show any structural homology to immunoglobulinproteins. Currently, two affilin scaffolds are employed, one of which isgamma crystalline, a human structural eye lens protein and the other is“ubiquitin” superfamily proteins. Both human scaffolds are very small,show high temperature stability and are almost resistant to pH changesand denaturing agents. This high stability is mainly due to the expandedbeta sheet structure of the proteins. Examples of gamma crystallinederived proteins are described in WO200104144 and examples of“ubiquitin-like” proteins are described in WO2004106368.

Protein epitope mimetics (PEM) are medium-sized, cyclic, peptide-likemolecules (MW 1-2 kDa) mimicking beta-hairpin secondary structures ofproteins, the major secondary structure involved in protein-proteininteractions.

In some embodiments, the Fabs are converted to silent IgG1 format bychanging the Fc region. For example, antibodies 6525-7910 in Table 1 canbe converted to silent IgG1 formate by substituting the “X” in the aminoacid sequences for the heavy chain with:

(SEQ ID NO: 293) CDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKand substituting the “X” in the amino acid sequence for the light chainwith: CS if the light chain is lambda, or C if the light chain is kappa.As used herein, a “silent IgG1” is an IgG1 Fc sequence in which theamino acid sequence has been altered to reduce Fc-mediated effectorfunctions (for example ADCC and/or CDC). Such an antibody will typicallyhave reduced binding to Fc receptors and/or C1q.

In some other embodiments, the Fabs are converted to IgG2 format. Forexample, antibodies 6525-7910 in Table 1 can be converted to IgG2 formatby substituting the constant sequence

(SEQ ID NO: 294) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEP KSXwith the constant sequence for the heavy chain of IgG2:

(SEQ ID NO: 295) ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKand substituting the “X” in the amino acid sequence for the light chainwith CS if the light chain is lambda, or C if the light chain is kappa.

Human or Humanized Antibodies

The present invention provides fully human antibodies that specificallybind to a C5 protein (e.g., human and/or cynomolgus C5). Compared to thechimeric or humanized antibodies, the human C5-binding antibodies of theinvention have further reduced antigenicity when administered to humansubjects.

The human C5-binding antibodies can be generated using methods that areknown in the art. For example, the humaneering technology used toconverting non-human antibodies into engineered human antibodies. U.S.Patent Publication No. 20050008625 describes an in vivo method forreplacing a nonhuman antibody variable region with a human variableregion in an antibody while maintaining the same or providing betterbinding characteristics relative to that of the nonhuman antibody. Themethod relies on epitope guided replacement of variable regions of anon-human reference antibody with a fully human antibody. The resultinghuman antibody is generally unrelated structurally to the referencenonhuman antibody, but binds to the same epitope on the same antigen asthe reference antibody. Briefly, the serial epitope-guidedcomplementarity replacement approach is enabled by setting up acompetition in cells between a “competitor” and a library of diversehybrids of the reference antibody (“lest antibodies”) for binding tolimiting amounts of antigen in the presence of a reporter system whichresponds to the binding of test antibody to antigen. The competitor canbe the reference antibody or derivative thereof such as a single-chainFv fragment. The competitor can also be a natural or artificial ligandof the antigen which binds to the same epitope as the referenceantibody. The only requirements of the competitor are that it binds tothe same epitope as the reference antibody, and that it competes withthe reference antibody for antigen binding. The test antibodies have oneantigen-binding V-region in common from the nonhuman reference antibody,and the other V-region selected at random from a diverse source such asa repertoire library of human antibodies. The common V-region from thereference antibody serves as a guide, positioning the test antibodies onthe same epitope on the antigen, and in the same orientation, so thatselection is biased toward the highest antigen-binding fidelity to thereference antibody.

Many types of reporter system can be used to detect desired interactionsbetween test antibodies and antigen. For example, complementing reporterfragments may be linked to antigen and test antibody, respectively, sothat reporter activation by fragment complementation only occurs whenthe test antibody binds to the antigen. When the test antibody- andantigen-reporter fragment fusions are co-expressed with a competitor,reporter activation becomes dependent on the ability of the testantibody to compete with the competitor, which is proportional to theaffinity of the test antibody for the antigen. Other reporter systemsthat can be used include the reactivator of an auto-inhibited reporterreactivation system (RAIR) as disclosed in U.S. patent application Ser.No. 10/208,730 (Publication No. 20030198971), or competitive activationsystem disclosed in U.S. patent application Ser. No. 10/076,845(Publication No. 20030157579).

With the serial epitope-guided complementarity replacement system,selection is made to identify cells expresses a single test antibodyalong with the competitor, antigen, and reporter components. In thesecells, each test antibody competes one-on-one with the competitor forbinding to a limiting amount of antigen. Activity of the reporter isproportional to the amount of antigen bound to the test antibody, whichin turn is proportional to the affinity of the test antibody for theantigen and the stability of the test antibody. Test antibodies areinitially selected on the basis of their activity relative to that ofthe reference antibody when expressed as the test antibody. The resultof the first round of selection is a set of “hybrid” antibodies, each ofwhich is comprised of the same non-human V-region from the referenceantibody and a human V-region from the library, and each of which bindsto the same epitope on the antigen as the reference antibody. One ofmore of the hybrid antibodies selected in the first round will have anaffinity for the antigen comparable to or higher than that of thereference antibody.

In the second V-region replacement step, the human V-regions selected inthe first step are used as guide for the selection of human replacementsfor the remaining non-human reference antibody V-region with a diverselibrary of cognate human V-regions. The hybrid antibodies selected inthe first round may also be used as competitors for the second round ofselection. The result of the second round of selection is a set of fullyhuman antibodies which differ structurally from the reference antibody,but which compete with the reference antibody for binding to the sameantigen. Some of the selected human antibodies bind to the same epitopeon the same antigen as the reference antibody. Among these selectedhuman antibodies, one or more binds to the same epitope with an affinitywhich is comparable to or higher than that of the reference antibody.

Using one of the mouse or chimeric C5-binding antibodies described aboveas the reference antibody, this method can be readily employed togenerate human antibodies that bind to human C5 with the same bindingspecificity and the same or better binding affinity. In addition, suchhuman C5-binding antibodies can also be commercially obtained fromcompanies which customarily produce human antibodies, e.g., KaloBios,Inc. (Mountain View, Calif.).

Camelid Antibodies

Antibody proteins obtained from members of the camel and dromedary(Camelus bactrianus and Calelus dromaderius) family including new worldmembers such as llama species (Lama paccos, Lama glama and Lama vicugna)have been characterized with respect to size, structural complexity andantigenicity for human subjects. Certain IgG antibodies from this familyof mammals as found in nature lack light chains, and are thusstructurally distinct from the typical four chain quaternary structurehaving two heavy and two light chains, for antibodies from otheranimals. See PCT/EP93/02214 (WO 94/04678 published 3 Mar. 1994).

A region of the camelid antibody which is the small single variabledomain identified as VHH can be obtained by genetic engineering to yielda small protein having high affinity for a target, resulting in a lowmolecular weight antibody-derived protein known as a “camelid nanobody”.See U.S. Pat. No. 5,759,808 issued Jun. 2, 1998; see also Stijlemans, B.et al., 2004 J Biol Chem 279: 1256-1261; Dumoulin, M. et al., 2003Nature 424: 783-788; Pleschberger, M. et al. 2003 Bioconjugate Chem 14:440-448; Cortez-Retamozo, V. et al. 2002 Int J Cancer 89: 456-62; andLauwereys, M. et al. 1998 EMBO J 17: 3512-3520. Engineered libraries ofcamelid antibodies and antibody fragments are commercially available,for example, from Ablynx, Ghent, Belgium. As with other antibodies ofnon-human origin, an amino acid sequence of a camelid antibody can bealtered recombinantly to obtain a sequence that more closely resembles ahuman sequence, i.e., the nanobody can be “humanized”. Thus the naturallow antigenicity of camelid antibodies to humans can be further reduced.

The camelid nanobody has a molecular weight approximately one-tenth thatof a human IgG molecule, and the protein has a physical diameter of onlya few nanometers. One consequence of the small size is the ability ofcamelid nanobodies to bind to antigenic sites that are functionallyinvisible to larger antibody proteins, i.e., camelid nanobodies areuseful as reagents detect antigens that are otherwise cryptic usingclassical immunological techniques, and as possible therapeutic agents.Thus yet another consequence of small size is that a camelid nanobodycan inhibit as a result of binding to a specific site in a groove ornarrow cleft of a target protein, and hence can serve in a capacity thatmore closely resembles the function of a classical low molecular weightdrug than that of a classical antibody.

The low molecular weight and compact size further result in camelidnanobodies being extremely thermostable, stable to extreme pH and toproteolytic digestion, and poorly antigenic. Another consequence is thatcamelid nanobodies readily move from the circulatory system intotissues, and even cross the blood-brain barrier and can treat disordersthat affect nervous tissue. Nanobodies can further facilitated drugtransport across the blood brain barrier. See U.S. patent application20040161738 published Aug. 19, 2004. These features combined with thelow antigenicity to humans indicate great therapeutic potential.Further, these molecules can be fully expressed in prokaryotic cellssuch as E. coli and are expressed as fusion proteins with bacteriophageand are functional.

Accordingly, a feature of the present invention is a camelid antibody ornanobody having high affinity for C5. In certain embodiments herein, thecamelid antibody or nanobody is naturally produced in the camelidanimal, i.e., is produced by the camelid following immunization with C5or a peptide fragment thereof, using techniques described herein forother antibodies. Alternatively, the C5-binding camelid nanobody isengineered, i.e., produced by selection for example from a library ofphage displaying appropriately mutagenized camelid nanobody proteinsusing panning procedures with C5 as a target as described in theexamples herein. Engineered nanobodies can further be customized bygenetic engineering to have a half life in a recipient subject of from45 minutes to two weeks. In a specific embodiment, the camelid antibodyor nanobody is obtained by grafting the CDRs sequences of the heavy orlight chain of the human antibodies of the invention into nanobody orsingle domain antibody framework sequences, as described for example inPCT/EP93/02214.

Bispecific Molecules and Multivalent Antibodies

In another aspect, the present invention features bispecific ormultispecific molecules comprising an C5-binding antibody, or a fragmentthereof, of the invention. An antibody of the invention, orantigen-binding regions thereof, can be derivatized or linked to anotherfunctional molecule, e.g., another peptide or protein (e.g., anotherantibody or ligand for a receptor) to generate a bispecific moleculethat binds to at least two different binding sites or target molecules.The antibody of the invention may in fact be derivatized or linked tomore than one other functional molecule to generate multi-specificmolecules that bind to more than two different binding sites and/ortarget molecules; such multi-specific molecules are also intended to beencompassed by the term “bispecific molecule” as used herein. To createa bispecific molecule of the invention, an antibody of the invention canbe functionally linked (e.g., by chemical coupling, genetic fusion,noncovalent association or otherwise) to one or more other bindingmolecules, such as another antibody, antibody fragment, peptide orbinding mimetic, such that a bispecific molecule results.

Accordingly, the present invention includes bispecific moleculescomprising at least one first binding specificity for C5 and a secondbinding specificity for a second target epitope. For example, the secondtarget epitope is another epitope of C5 different from the first targetepitope.

Additionally, for the invention in which the bispecific molecule ismulti-specific, the molecule can further include a third bindingspecificity, in addition to the first and second target epitope.

In one embodiment, the bispecific molecules of the invention comprise asa binding specificity at least one antibody, or an antibody fragmentthereof, including, e.g., an Fab, Fab′, F(ab′)2, Fv, or a single chainFv. The antibody may also be a light chain or heavy chain dimer, or anyminimal fragment thereof such as a Fv or a single chain construct asdescribed in Ladner et al. U.S. Pat. No. 4,946,778.

Diabodies are bivalent, bispecific molecules in which VH and VL domainsare expressed on a single polypeptide chain, connected by a linker thatis too short to allow for pairing between the two domains on the samechain. The VH and VL domains pair with complementary domains of anotherchain, thereby creating two antigen binding sites (see e.g., Holliger etal., 1993 Proc. Natl. Acad. Sci. USA 90:6444-6448; Poijak et al., 1994

Structure 2:1121-1123). Diabodies can be produced by expressing twopolypeptide chains with either the structure VHA-VLB and VHB-VLA (VH-VLconfiguration), or VLA-VHB and VLB-VHA (VL-VH configuration) within thesame cell. Most of them can be expressed in soluble form in bacteria.Single chain diabodies (scDb) are produced by connecting the twodiabody-forming polypeptide chains with linker of approximately 15 aminoacid residues (see Holliger and Winter, 1997 Cancer Immunol.Immunother., 45(3-4):128-30; Wu et al., 1996 Immunotechnology,2(1):21-36). scDb can be expressed in bacteria in soluble, activemonomeric form (see Holliger and Winter, 1997 Cancer Immunol.Immunother., 45(34): 128-30; Wu et al., 1996 Immunotechnology,2(1):21-36; Pluckthun and Pack, 1997 Immunotechnology, 3(2): 83-105;Ridgway et al., 1996 Protein Eng., 9(7):617-21). A diabody can be fusedto Fc to generate a “di-diabody” (see Lu et al., 2004 J. Biol. Chem.,279(4):2856-65).

Other antibodies which can be employed in the bispecific molecules ofthe invention are murine, chimeric and humanized monoclonal antibodies.

The bispecific molecules of the present invention can be prepared byconjugating the constituent binding specificities, using methods knownin the art. For example, each binding specificity of the bispecificmolecule can be generated separately and then conjugated to one another.When the binding specificities are proteins or peptides, a variety ofcoupling or cross-linking agents can be used for covalent conjugation.Examples of cross-linking agents include protein A, carbodiimide,N-succinimidyl-5-acetyl-thioacetate (SATA),5,5′-dithiobis(2-nitrobenzoic acid) (DTNB), o-phenylenedimaleimide(oPDM), N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), andsulfosuccinimidyl 4-(N-maleimidomethyl)cyclohaxane-1-carboxylate(sulfo-SMCC) (see e.g., Karpovsky et al., 1984 J. Exp. Med. 160:1686;Liu, M A et al., 1985 Proc. Natl. Acad. Sci. USA 82:8648). Other methodsinclude those described in Paulus, 1985 Behring Ins. Mitt. No. 78,118-132; Brennan et al., 1985 Science 229:81-83), and Glennie et al.,1987 J. Immunol. 139: 2367-2375). Conjugating agents are SATA andsulfo-SMCC, both available from Pierce Chemical Co. (Rockford, Ill.).

When the binding specificities are antibodies, they can be conjugated bysulfhydryl bonding of the C-terminus hinge regions of the two heavychains. In a particularly embodiment, the hinge region is modified tocontain an odd number of sulfhydryl residues, for example one, prior toconjugation.

Alternatively, both binding specificities can be encoded in the samevector and expressed and assembled in the same host cell. This method isparticularly useful where the bispecific molecule is a mAb×mAb, mAb×Fab,Fab×F(ab′)2 or ligand×Fab fusion protein. A bispecific molecule of theinvention can be a single chain molecule comprising one single chainantibody and a binding determinant, or a single chain bispecificmolecule comprising two binding determinants. Bispecific molecules maycomprise at least two single chain molecules. Methods for preparingbispecific molecules are described for example in U.S. Pat. No.5,260,203; U.S. Pat. No. 5,455,030; U.S. Pat. No. 4,881,175; U.S. Pat.No. 5,132,405; U.S. Pat. No. 5,091,513; U.S. Pat. No. 5,476,786; U.S.Pat. No. 5,013,653; U.S. Pat. No. 5,258,498; and U.S. Pat. No.5,482,858.

Binding of the bispecific molecules to their specific targets can beconfirmed by, for example, enzyme-linked immunosorbent assay (ELISA),radioimmunoassay (REA), FACS analysis, bioassay (e.g., growthinhibition), or Western Blot assay. Each of these assays generallydetects the presence of protein-antibody complexes of particularinterest by employing a labeled reagent (e.g., an antibody) specific forthe complex of interest.

In another aspect, the present invention provides multivalent compoundscomprising at least two identical or different antigen-binding portionsof the antibodies of the invention binding to C5. The antigen-bindingportions can be linked together via protein fusion or covalent or noncovalent linkage. Alternatively, methods of linkage has been describedfor the bispecific molecules. Tetravalent compounds can be obtained forexample by cross-linking antibodies of the antibodies of the inventionwith an antibody that binds to the constant regions of the antibodies ofthe invention, for example the Fc or hinge region.

Trimerizing domain are described for example in Borean patent EP 1 012280B1. Pentamerizing modules are described for example inPCT/EP97/05897.

Antibodies with Extended Half Life

The present invention provides for antibodies that specifically bind toC5 protein which have an extended half-life in vivo.

Many factors may affect a protein's half life in vivo. For examples,kidney filtration, metabolism in the liver, degradation by proteolyticenzymes (proteases), and immunogenic responses (e.g., proteinneutralization by antibodies and uptake by macrophages and dentriticcells). A variety of strategies can be used to extend the half life ofthe antibodies of the present invention. For example, by chemicallinkage to polyethyleneglycol (PEG), reCODE PEG, antibody scaffold,polysialic acid (PSA), hydroxyethyl starch (HES), albumin-bindingligands, and carbohydrate shields; by genetic fusion to proteins bindingto serum proteins, such as albumin, IgG, FcRn, and transferring; bycoupling (genetically or chemically) to other binding moieties that bindto serum proteins, such as nanobodies, Fabs, DARPins, avimers,affibodies, and anticalins; by genetic fusion to rPEG, albumin, domainof albumin, albumin-binding proteins, and Fc; or by incorporation intonancarriers, slow release formulations, or medical devices.

To prolong the serum circulation of antibodies in vivo, inert polymermolecules such as high molecular weight PEG can be attached to theantibodies or a fragment thereof with or without a multifunctionallinker either through site-specific conjugation of the PEG to the N- orC-terminus of the antibodies or via epsilon-amino groups present onlysine residues. To pegylate an antibody, the antibody, or fragmentthereof, typically is reacted with polyethylene glycol (PEG), such as areactive ester or aldehyde derivative of PEG, under conditions in whichone or more PEG groups become attached to the antibody or antibodyfragment. The pegylation can be carried out by an acylation reaction oran alkylation reaction with a reactive PEG molecule (or an analogousreactive water-soluble polymer). As used herein, the term “polyethyleneglycol” is intended to encompass any of the forms of PEG that have beenused to derivatize other proteins, such as mono (C1-C10)alkoxy- oraryloxy-polyethylene glycol or polyethylene glycol-maleimide. In certainembodiments, the antibody to be pegylated is an aglycosylated antibody.Linear or branched polymer derivatization that results in minimal lossof biological activity will be used. The degree of conjugation can beclosely monitored by SDS-PAGE and mass spectrometry to ensure properconjugation of PEG molecules to the antibodies. Unreacted PEG can beseparated from antibody-PEG conjugates by size-exclusion or byion-exchange chromatography. PEG-derivatized antibodies can be testedfor binding activity as well as for in vivo efficacy using methodswell-known to those of skill in the art, for example, by immunoassaysdescribed herein. Methods for pegylating proteins are known in the artand can be applied to the antibodies of the invention. See for example,EP 0 154 316 by Nishimura et al. and EP 0 401 384 by Ishikawa et al.

Other modified pegylation technologies include reconstituting chemicallyorthogonal directed engineering technology (ReCODE PEG), whichincorporates chemically specified side chains into biosynthetic proteinsvia a reconstituted system that includes tRNA synthetase and tRNA. Thistechnology enables incorporation of more than 30 new amino acids intobiosynthetic proteins in E. coli, yeast, and mammalian cells. The tRNAincorporates a normative amino acid any place an amber codon ispositioned, converting the amber from a stop codon to one that signalsincorporation of the chemically specified amino acid.

Recombinant pegylation technology (rPEG) can also be used for serumhalflife extension. This technology involves genetically fusing a300-600 amino acid unstructured protein tail to an existingpharmaceutical protein. Because the apparent molecular weight of such anunstructured protein chain is about 15-fold larger than its actualmolecular weight, the serum halflife of the protein is greatlyincreased. In contrast to traditional PEGylation, which requireschemical conjugation and repurification, the manufacturing process isgreatly simplified and the product is homogeneous.

Polysialytion is another technology, which uses the natural polymerpolysialic acid (PSA) to prolong the active life and improve thestability of therapeutic peptides and proteins. PSA is a polymer ofsialic acid (a sugar). When used for protein and therapeutic peptidedrug delivery, polysialic acid provides a protective microenvironment onconjugation. This increases the active life of the therapeutic proteinin the circulation and prevents it from being recognized by the immunesystem. The PSA polymer is naturally found in the human body. It wasadopted by certain bacteria which evolved over millions of years to coattheir walls with it. These naturally polysialylated bacteria were thenable, by virtue of molecular mimicry, to foil the body's defense system.PSA, nature's ultimate stealth technology, can be easily produced fromsuch bacteria in large quantities and with predetermined physicalcharacteristics. Bacterial PSA is completely non-immunogenic, even whencoupled to proteins, as it is chemically identical to PSA in the humanbody.

Another technology include the use of hydroxyethyl starch (“HES”)derivatives linked to antibodies. HES is a modified natural polymerderived from waxy maize starch and can be metabolized by the body'senzymes. HES solutions are usually administered to substitute deficientblood volume and to improve the rheological properties of the blood.Hesylation of an antibody enables the prolongation of the circulationhalf-life by increasing the stability of the molecule, as well as byreducing renal clearance, resulting in an increased biological activity.By varying different parameters, such as the molecular weight of HES, awide range of HES antibody conjugates can be customized.

Antibodies having an increased half-life in vivo can also be generatedintroducing one or more amino acid modifications (i.e., substitutions,insertions or deletions) into an IgG constant domain, or FcRn bindingfragment thereof (preferably a Fc or hinge Fc domain fragment). See,e.g., International Publication No. WO 98/23289; InternationalPublication No. WO 97/34631; and U.S. Pat. No. 6,277,375.

Further, antibodies can be conjugated to albumin in order to make theantibody or antibody fragment more stable in vivo or have a longer halflife in vivo. The techniques are well-known in the art, see, e.g.,International Publication Nos. WO 93/15199, WO 93/15200, and WO01/77137; and European Patent No. EP 413,622.

The strategies for increasing half life is especially useful innanobodies, fibronectin-based binders, and other antibodies or proteinsfor which increased in vivo half life is desired.

Antibody Conjugates

The present invention provides antibodies or fragments thereof thatspecifically bind to a C5 protein recombinantly fused or chemicallyconjugated (including both covalent and non-covalent conjugations) to aheterologous protein or polypeptide (or fragment thereof, preferably toa polypeptide of at least 10, at least 20, at least 30, at least 40, atleast 50, at least 60, at least 70, at least 80, at least 90 or at least100 amino acids) to generate fusion proteins. In particular, theinvention provides fusion proteins comprising an antigen-bindingfragment of an antibody described herein (e.g., a Fab fragment, Fdfragment, Fv fragment, F(ab)₂ fragment, a VH domain, a VH CDR, a VLdomain or a VL CDR) and a heterologous protein, polypeptide, or peptide.Methods for fusing or conjugating proteins, polypeptides, or peptides toan antibody or an antibody fragment are known in the art. See, e.g.,U.S. Pat. Nos. 5,336,603, 5,622,929, 5,359,046, 5,349,053, 5,447,851,and 5,112,946; European Patent Nos. EP 307,434 and EP 367,166;International Publication Nos. WO 96/04388 and WO 91/06570; Ashkenazi etal., 1991, Proc. Natl. Acad. Sci. USA 88: 10535-10539; Zheng et al.,1995, J. Immunol. 154:5590-5600; and Vil et al., 1992, Proc. Natl. Acad.Sci. USA 89:11337-11341.

Additional fusion proteins may be generated through the techniques ofgene-shuffling, motif-shuffling, exon-shuffling, and/or codon-shuffling(collectively referred to as “DNA shuffling”). DNA shuffling may beemployed to alter the activities of antibodies of the invention orfragments thereof (e.g., antibodies or fragments thereof with higheraffinities and lower dissociation rates). See, generally, U.S. Pat. Nos.5,605,793, 5,811,238, 5,830,721, 5,834,252, and 5,837,458; Patten etal., 1997, Curr. Opinion Biotechnol. 8:724-33; Harayama, 1998, TrendsBiotechnol. 16(2):76-82; Hansson, et al., 1999, J. Mol. Biol.287:265-76; and Lorenzo and Blasco, 1998, Biotechniques 24(2):308-313(each of these patents and publications are hereby incorporated byreference in its entirety). Antibodies or fragments thereof, or theencoded antibodies or fragments thereof, may be altered by beingsubjected to random mutagenesis by error-prone PCR, random nucleotideinsertion or other methods prior to recombination. A polynucleotideencoding an antibody or fragment thereof that specifically binds to a C5protein may be recombined with one or more components, motifs, sections,parts, domains, fragments, etc. of one or more heterologous molecules.

Moreover, the antibodies or fragments thereof can be fused to markersequences, such as a peptide to facilitate purification. In preferredembodiments, the marker amino acid sequence is a hexa-histidine peptide,such as the tag provided in a pQE vector (QIAGEN, Inc., 9259 EtonAvenue, Chatsworth, Calif., 91311), among others, many of which arecommercially available. As described in Gentz et al., 1989, Proc. Natl.Acad. Sci. USA 86:821-824, for instance, hexa-histidine provides forconvenient purification of the fusion protein. Other peptide tags usefulfor purification include, but are not limited to, the hemagglutinin(“HA”) tag, which corresponds to an epitope derived from the influenzahemagglutinin protein (Wilson et al., 1984, Cell 37:767), and the “flag”tag.

In other embodiments, antibodies of the present invention or fragmentsthereof conjugated to a diagnostic or detectable agent. Such antibodiescan be useful for monitoring or prognosing the onset, development,progression and/or severity of a disease or disorder as part of aclinical testing procedure, such as determining the efficacy of aparticular therapy. Such diagnosis and detection can accomplished bycoupling the antibody to detectable substances including, but notlimited to, various enzymes, such as, but not limited to, horseradishperoxidase, alkaline phosphatase, beta-galactosidase, oracetylcholinesterase; prosthetic groups, such as, but not limited to,streptavidin/biotin and avidin/biotin; fluorescent materials, such as,but not limited to, umbelliferone, fluorescein, fluoresceinisothiocynate, rhodamine, dichlorotriazinylamine fluorescein, dansylchloride or phycoerythrin; luminescent materials, such as, but notlimited to, luminol; bioluminescent materials, such as but not limitedto, luciferase, luciferin, and aequorin; radioactive materials, such as,but not limited to, iodine (131I, 125I, 123I, and 121I), carbon (14C),sulfur (35S), tritium (3H), indium (115In, 113In, 112In, and 111In),technetium (99Tc), thallium (201Ti), gallium (68Ga, 67Ga), palladium(103Pd), molybdenum (99Mo), xenon (133Xe), fluorine (18F), 153Sm, 177Lu,159Gd, 149 Pm, 140La, 175Yb, 166Ho, 90Y, 47Sc, 186Re, 188Re, 142Pr,105Rh, 97Ru, 68Ge, 57Co, 65Zn, 85Sr, 32P, 153Gd, 169Yb, 51Cr, 54Mn,75Se, 113Sn, and 117Tin; and positron emitting metals using variouspositron emission tomographies, and nonradioactive paramagnetic metalions.

The present invention further encompasses uses of antibodies orfragments thereof conjugated to a therapeutic moiety. An antibody orfragment thereof may be conjugated to a therapeutic moiety such as acytotoxin, e.g., a cytostatic or cytocidal agent, a therapeutic agent ora radioactive metal ion, e.g., alpha-emitters. A cytotoxin or cytotoxicagent includes any agent that is detrimental to cells.

Further, an antibody or fragment thereof may be conjugated to atherapeutic moiety or drug moiety that modifies a given biologicalresponse. Therapeutic moieties or drug moieties are not to be construedas limited to classical chemical therapeutic agents. For example, thedrug moiety may be a protein, peptide, or polypeptide possessing adesired biological activity. Such proteins may include, for example, atoxin such as abrin, ricin A, pseudomonas exotoxin, cholera toxin, ordiphtheria toxin; a protein such as tumor necrosis factor, α-interferon,β-interferon, nerve growth factor, platelet derived growth factor,tissue plasminogen activator, an apoptotic agent, an anti-angiogenicagent; or, a biological response modifier such as, for example, alymphokine.

Moreover, an antibody can be conjugated to therapeutic moieties such asa radioactive metal ion, such as alpha-emitters such as 213Bi ormacrocyclic chelators useful for conjugating radiometal ions, includingbut not limited to, 131In, 131LU, 131Y, 131Ho, 131Sm, to polypeptides.In certain embodiments, the macrocyclic chelator is1,4,7,10-tetraazacyclododecane-N,N′,N″,N′″-tetraacetic acid (DOTA) whichcan be attached to the antibody via a linker molecule. Such linkermolecules are commonly known in the art and described in Denardo et al.,1998, Clin Cancer Res. 4(10):2483-90; Peterson et al., 1999, Bioconjug.Chem. 10(4):553-7; and Zimmerman et al., 1999, Nucl. Med. Biol.26(8):943-50, each incorporated by reference in their entireties.

Techniques for conjugating therapeutic moieties to antibodies are wellknown, see, e.g., Amon et al., “Monoclonal Antibodies ForImmunotargeting Of Drugs In Cancer Therapy”, in Monoclonal AntibodiesAnd Cancer Therapy, Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss,Inc. 1985); Hellstrom et al., “Antibodies For Drug Delivery”, inControlled Drug Delivery (2nd Ed.), Robinson et al. (eds.), pp. 623-53(Marcel Dekker, Inc. 1987); Thorpe, “Antibody Carriers Of CytotoxicAgents In Cancer Therapy: A Review”, in Monoclonal Antibodies 84:Biological And Clinical Applications, Pinchera et al. (eds.), pp.475-506 (1985); “Analysis, Results, And Future Prospective Of TheTherapeutic Use Of Radiolabeled Antibody In Cancer Therapy”, inMonoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al.(eds.), pp. 303-16 (Academic Press 1985), and Thorpe et al., 1982,Immunol. Rev. 62:119-58.

Antibodies may also be attached to solid supports, which areparticularly useful for immunoassays or purification of the targetantigen. Such solid supports include, but are not limited to, glass,cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride orpolypropylene.

5.2. METHODS OF PRODUCING ANTIBODIES OF THE INVENTION 5.2.1. NucleicAcids Encoding the Antibodies

The invention provides substantially purified nucleic acid moleculeswhich encode polypeptides comprising segments or domains of theC5-binding antibody chains described above. Some of the nucleic acids ofthe invention comprise the nucleotide sequence encoding the heavy chainvariable region shown in SEQ ID NO: 7, 23, 39, 51, 67, 79, 96, 108, 114,121, 137, 151, 165, 179, 187, 201, 210, 218, 227, 241, 253, 257, 273,277, or 281, and/or the nucleotide sequence encoding the light chainvariable region shown in SEQ ID NO: 8, 24, 40, 52, 68, 80, 90, 102, 122,138, 152, 166, 180, 188, 202, 211, 219, 228, 242, 261, 265, 269, 285, or289. In a specific embodiment, the nucleic acid molecules are thoseidentified in Table 1. Some other nucleic acid molecules of theinvention comprise nucleotide sequences that are substantially identical(e.g., at least 65, 80%, 95%, or 99%) to the nucleotide sequences ofthose identified in Table 1. When expressed from appropriate expressionvectors, polypeptides encoded by these polynucleotides are capable ofexhibiting C5 antigen binding capacity.

Also provided in the invention are polynucleotides which encode at leastone CDR region and usually all three CDR regions from the heavy or lightchain of the C5-binding antibody set forth above. Some otherpolynucleotides encode all or substantially all of the variable regionsequence of the heavy chain and/or the light chain of the C5-bindingantibody set forth above. Because of the degeneracy of the code, avariety of nucleic acid sequences will encode each of the immunoglobulinamino acid sequences.

The nucleic acid molecules of the invention can encode both a variableregion and a constant region of the antibody. Some of nucleic acidsequences of the invention comprise nucleotides encoding a mature heavychain variable region sequence that is substantially identical (e.g., atleast 80%, 90%, or 99%) to the mature heavy chain variable regionsequence set forth in SEQ ID NO: 7, 23, 39, 51, 67, 79, 96, 108, 114,121, 137, 151, 165, 179, 187, 201, 210, 218, 227, 241, 253, 257, 273,277, or 281. Some other nucleic acid sequences comprising nucleotideencoding a mature light chain variable region sequence that issubstantially identical (e.g., at least 80%, 90%, or 99%) to the maturelight chain variable region sequence set forth in SEQ ID NO: 8, 24, 40,52, 68, 80, 90, 102, 122, 138, 152, 166, 180, 188, 202, 211, 219, 228,242, 261, 265, 269, 285, or 289.

The polynucleotide sequences can be produced by de novo solid-phase DNAsynthesis or by PCR mutagenesis of an existing sequence (e.g., sequencesas described in the Examples below) encoding an C5-binding antibody orits binding fragment. Direct chemical synthesis of nucleic acids can beaccomplished by methods known in the art, such as the phosphotriestermethod of Narang et al., 1979, Meth. Enzymol. 68:90; the phosphodiestermethod of Brown et al., Meth. Enzymol. 68:109, 1979; thediethylphosphoramidite method of Beaucage et al., Tetra. Lett., 22:1859,1981; and the solid support method of U.S. Pat. No. 4,458,066.Introducing mutations to a polynucleotide sequence by PCR can beperformed as described in, e.g., PCR Technology: Principles andApplications for DNA Amplification, H. A. Erlich (Ed.), Freeman Press,NY, N.Y., 1992; PCR Protocols: A Guide to Methods and Applications,Innis et al. (Ed.), Academic Press, San Diego, Calif., 1990; Mattila etal., Nucleic Acids Res. 19:967, 1991; and Eckert et al., PCR Methods andApplications 1:17, 1991.

Also provided in the invention are expression vectors and host cells forproducing the C5-binding antibodies described above. Various expressionvectors can be employed to express the polynucleotides encoding theC5-binding antibody chains or binding fragments. Both viral-based andnonviral expression vectors can be used to produce the antibodies in amammalian host cell. Nonviral vectors and systems include plasmids,episomal vectors, typically with an expression cassette for expressing aprotein or RNA, and human artificial chromosomes (see, e.g., Harringtonet al., Nat Genet. 15:345, 1997). For example, nonviral vectors usefulfor expression of the C5-binding polynucleotides and polypeptides inmammalian (e.g., human) cells include pThioHis A, B & C, pcDNA3.1/His,pEBVHis A, B & C, (Invitrogen, San Diego, Calif.), MPSV vectors, andnumerous other vectors known in the art for expressing other proteins.Useful viral vectors include vectors based on retroviruses,adenoviruses, adenoassociated viruses, herpes viruses, vectors based onSV40, papilloma virus, HBP Epstein Barr virus, vaccinia virus vectorsand Semliki Forest virus (SFV). See, Brent et al., supra; Smith, Annu.Rev. Microbiol. 49:807, 1995; and Rosenfeld et al., Cell 68:143, 1992.

The choice of expression vector depends on the intended host cells inwhich the vector is to be expressed. Typically, the expression vectorscontain a promoter and other regulatory sequences (e.g., enhancers) thatare operably linked to the polynucleotides encoding an C5-bindingantibody chain or fragment. In some embodiments, an inducible promoteris employed to prevent expression of inserted sequences except underinducing conditions. Inducible promoters include, e.g., arabinose, lacZ,metallothionein promoter or a heat shock promoter. Cultures oftransformed organisms can be expanded under noninducing conditionswithout biasing the population for coding sequences whose expressionproducts are better tolerated by the host cells. In addition topromoters, other regulatory elements may also be required or desired forefficient expression of an C5-binding antibody chain or fragment. Theseelements typically include an ATG initiation codon and adjacent ribosomebinding site or other sequences. In addition, the efficiency ofexpression may be enhanced by the inclusion of enhancers appropriate tothe cell system in use (see, e.g., Scharf et al., Results Probl. CellDiffer. 20:125, 1994; and Bittner et al., Meth. Enzymol., 153:516,1987). For example, the SV40 enhancer or CMV enhancer may be used toincrease expression in mammalian host cells.

The expression vectors may also provide a secretion signal sequenceposition to form a fusion protein with polypeptides encoded by insertedC5-binding antibody sequences. More often, the inserted C5-bindingantibody sequences are linked to a signal sequences before inclusion inthe vector. Vectors to be used to receive sequences encoding C5-bindingantibody light and heavy chain variable domains sometimes also encodeconstant regions or parts thereof. Such vectors allow expression of thevariable regions as fusion proteins with the constant regions therebyleading to production of intact antibodies or fragments thereof.Typically, such constant regions are human.

The host cells for harboring and expressing the C5-binding antibodychains can be either prokaryotic or eukaryotic. E. coli is oneprokaryotic host useful for cloning and expressing the polynucleotidesof the present invention. Other microbial hosts suitable for use includebacilli, such as Bacillus subtilis, and other enterobacteriaceae, suchas Salmonella, Serratia, and various Pseudomonas species. In theseprokaryotic hosts, one can also make expression vectors, which typicallycontain expression control sequences compatible with the host cell(e.g., an origin of replication). In addition, any number of a varietyof well-known promoters will be present, such as the lactose promotersystem, a tryptophan (trp) promoter system, a beta-lactamase promotersystem, or a promoter system from phage lambda. The promoters typicallycontrol expression, optionally with an operator sequence, and haveribosome binding site sequences and the like, for initiating andcompleting transcription and translation. Other microbes, such as yeast,can also be employed to express C5-binding polypeptides of theinvention. Insect cells in combination with baculovirus vectors can alsobe used.

In some preferred embodiments, mammalian host cells are used to expressand produce the C5-binding polypeptides of the present invention. Forexample, they can be either a hybridoma cell line expressing endogenousimmunoglobulin genes (e.g., the 1D6.C9 myeloma hybridoma clone asdescribed in the Examples) or a mammalian cell line harboring anexogenous expression vector (e.g., the SP2/0 myeloma cells exemplifiedbelow). These include any normal mortal or normal or abnormal immortalanimal or human cell. For example, a number of suitable host cell linescapable of secreting intact immunoglobulins have been developedincluding the CHO cell lines, various Cos cell lines, HeLa cells,myeloma cell lines, transformed B-cells and hybridomas. The use ofmammalian tissue cell culture to express polypeptides is discussedgenerally in, e.g., Winnacker, FROM GENES TO CLONES, VCH Publishers,N.Y., N.Y., 1987. Expression vectors for mammalian host cells caninclude expression control sequences, such as an origin of replication,a promoter, and an enhancer (see, e.g., Queen, et al., Immunol. Rev.89:49-68, 1986), and necessary processing information sites, such asribosome binding sites, RNA splice sites, polyadenylation sites, andtranscriptional terminator sequences. These expression vectors usuallycontain promoters derived from mammalian genes or from mammalianviruses. Suitable promoters may be constitutive, cell type-specific,stage-specific, and/or modulatable or regulatable. Useful promotersinclude, but are not limited to, the metallothionein promoter, theconstitutive adenovirus major late promoter, the dexamethasone-inducibleMMTV promoter, the SV40 promoter, the MRP poIIII promoter, theconstitutive MPSV promoter, the tetracycline-inducible CMV promoter(such as the human immediate-early CMV promoter), the constitutive CMVpromoter, and promoter-enhancer combinations known in the art.

Methods for introducing expression vectors containing the polynucleotidesequences of interest vary depending on the type of cellular host. Forexample, calcium chloride transfection is commonly utilized forprokaryotic cells, whereas calcium phosphate treatment orelectroporation may be used for other cellular hosts. (See generallySambrook, et al., supra). Other methods include, e.g., electroporation,calcium phosphate treatment, liposome-mediated transformation, injectionand microinjection, ballistic methods, virosomes, immunoliposomes,polycation:nucleic acid conjugates, naked DNA, artificial virions,fusion to the herpes virus structural protein VP22 (Elliot and O'Hare,Cell 88:223, 1997), agent-enhanced uptake of DNA, and ex vivotransduction. For long-term, high-yield production of recombinantproteins, stable expression will often be desired. For example, celllines which stably express C5-binding antibody chains or bindingfragments can be prepared using expression vectors of the inventionwhich contain viral origins of replication or endogenous expressionelements and a selectable marker gene. Following the introduction of thevector, cells may be allowed to grow for 1-2 days in an enriched mediabefore they are switched to selective media. The purpose of theselectable marker is to confer resistance to selection, and its presenceallows growth of cells which successfully express the introducedsequences in selective media. Resistant, stably transfected cells can beproliferated using tissue culture techniques appropriate to the celltype.

5.2.2. Generation of Monoclonal Antibodies of the Invention

Monoclonal antibodies (mAbs) can be produced by a variety of techniques,including conventional monoclonal antibody methodology e.g., thestandard somatic cell hybridization technique of Kohler and Milstein,1975 Nature 256: 495. Many techniques for producing monoclonal antibodycan be employed e.g., viral or oncogenic transformation of Blymphocytes.

An animal system for preparing hybridomas is the murine system.Hybridoma production in the mouse is a well established procedure.Immunization protocols and techniques for isolation of immunizedsplenocytes for fusion are known in the art. Fusion partners (e.g.,murine myeloma cells) and fusion procedures are also known.

Chimeric or humanized antibodies of the present invention can beprepared based on the sequence of a murine monoclonal antibody preparedas described above. DNA encoding the heavy and light chainimmunoglobulins can be obtained from the murine hybridoma of interestand engineered to contain non-murine (e.g., human) immunoglobulinsequences using standard molecular biology techniques. For example, tocreate a chimeric antibody, the murine variable regions can be linked tohuman constant regions using methods known in the art (see e.g., U.S.Pat. No. 4,816,567 to Cabilly et al.). To create a humanized antibody,the murine CDR regions can be inserted into a human framework usingmethods known in the art. See e.g., U.S. Pat. No. 5,225,539 to Winter,and U.S. Pat. Nos. 5,530,101; 5,585,089; 5,693,762 and 6180370 to Queenet al.

In a certain embodiment, the antibodies of the invention are humanmonoclonal antibodies. Such human monoclonal antibodies directed againstC5 can be generated using transgenic or transchromosomic mice carryingparts of the human immune system rather than the mouse system. Thesetransgenic and transchromosomic mice include mice referred to herein asHuMAb mice and KM mice, respectively, and are collectively referred toherein as “human Ig mice.”

The HuMAb Mouse® (Medarex, Inc.) contains human immunoglobulin geneminiloci that encode un-rearranged human heavy (μ and γ) and κ lightchain immunoglobulin sequences, together with targeted mutations thatinactivate the endogenous μ and κ chain loci (see e.g., Lonberg, et al.,1994 Nature 368(6474): 856-859). Accordingly, the mice exhibit reducedexpression of mouse IgM or K, and in response to immunization, theintroduced human heavy and light chain transgenes undergo classswitching and somatic mutation to generate high affinity human IgGκmonoclonal (Lonberg, N. et al., 1994 supra; reviewed in Lonberg, N.,1994 Handbook of Experimental Pharmacology 113:49-101; Lonberg, N. andHuszar, D., 1995 Intern. Rev. Immunol. 13: 65-93, and Harding, F. andLonberg, N., 1995 Ann. N.Y. Acad. Sci. 764:536-546). The preparation anduse of HuMAb mice, and the genomic modifications carried by such mice,is further described in Taylor, L. et al., 1992 Nucleic Acids Research20:6287-6295; Chen, J. et al., 1993 International Immunology 5: 647-656;Tuaillon et al., 1993 Proc. Natl. Acad. Sci. USA 94:3720-3724; Choi etal., 1993 Nature Genetics 4:117-123; Chen, J. et al., 1993 EMBO J. 12:821-830; Tuaillon et al., 1994 J. Immunol. 152:2912-2920; Taylor, L. etal., 1994 International Immunology 579-591; and Fishwild, D. et al.,1996 Nature Biotechnology 14: 845-851, the contents of all of which arehereby specifically incorporated by reference in their entirety. Seefurther, U.S. Pat. Nos. 5,545,806; 5,569,825; 5,625,126; 5,633,425;5,789,650; 5,877,397; 5,661,016; 5,814,318; 5,874,299; and 5,770,429;all to Lonberg and Kay; U.S. Pat. No. 5,545,807 to Surani et al.; PCTPublication Nos. WO 92103918, WO 93/12227, WO 94/25585, WO 97113852, WO98/24884 and WO 99/45962, all to Lonberg and Kay; and PCT PublicationNo. WO 01/14424 to Korman et al.

In another embodiment, human antibodies of the invention can be raisedusing a mouse that carries human immunoglobulin sequences on transgenesand transchomosomes such as a mouse that carries a human heavy chaintransgene and a human light chain transchromosome. Such mice, referredto herein as “KM mice”, are described in detail in PCT Publication WO02/43478 to Ishida et al.

Still further, alternative transgenic animal systems expressing humanimmunoglobulin genes are available in the art and can be used to raiseC5-binding antibodies of the invention. For example, an alternativetransgenic system referred to as the Xenomouse (Abgenix, Inc.) can beused. Such mice are described in, e.g., U.S. Pat. Nos. 5,939,598;6,075,181; 6,114,598; 6,150,584 and 6,162,963 to Kucherlapati et al.

Moreover, alternative transchromosomic animal systems expressing humanimmunoglobulin genes are available in the art and can be used to raiseC5-binding antibodies of the invention. For example, mice carrying botha human heavy chain transchromosome and a human light chaintranschromosome, referred to as “TC mice” can be used; such mice aredescribed in Tomizuka et al., 2000 Proc. Natl. Acad. Sci. USA97:722-727. Furthermore, cows carrying human heavy and light chaintranschromosomes have been described in the art (Kuroiwa et al., 2002Nature Biotechnology 20:889-894) and can be used to raise C5-bindingantibodies of the invention.

Human monoclonal antibodies of the invention can also be prepared usingphage display methods for screening libraries of human immunoglobulingenes. Such phage display methods for isolating human antibodies areestablished in the art or described in the examples below. See forexample: U.S. Pat. Nos. 5,223,409; 5,403,484; and 5,571,698 to Ladner etal; U.S. Pat. Nos. 5,427,908 and 5,580,717 to Dower et al; U.S. Pat.Nos. 5,969,108 and 6,172,197 to McCafferty et al; and U.S. Pat. Nos.5,885,793; 6,521,404; 6,544,731; 6,555,313; 6,582,915 and 6,593,081 toGriffiths et al.

Human monoclonal antibodies of the invention can also be prepared usingSCID mice into which human immune cells have been reconstituted suchthat a human antibody response can be generated upon immunization. Suchmice are described in, for example, U.S. Pat. Nos. 5,476,996 and5,698,767 to Wilson et al.

5.2.3. Framework or Fc Engineering

Engineered antibodies of the invention include those in whichmodifications have been made to framework residues within VH and/or VL,e.g. to improve the properties of the antibody. Typically such frameworkmodifications are made to decrease the immunogenicity of the antibody.For example, one approach is to “backmutate” one or more frameworkresidues to the corresponding germline sequence. More specifically, anantibody that has undergone somatic mutation may contain frameworkresidues that differ from the germline sequence from which the antibodyis derived. Such residues can be identified by comparing the antibodyframework sequences to the germline sequences from which the antibody isderived. To return the framework region sequences to their germlineconfiguration, the somatic mutations can be “backmutated” to thegermline sequence by, for example, site-directed mutagenesis. Such“backmutated” antibodies are also intended to be encompassed by theinvention.

Another type of framework modification involves mutating one or moreresidues within the framework region, or even within one or more CDRregions, to remove T cell-epitopes to thereby reduce the potentialimmunogenicity of the antibody. This approach is also referred to as“deimmunization” and is described in further detail in U.S. PatentPublication No. 20030153043 by Carr et al.

In addition or alternative to modifications made within the framework orCDR regions, antibodies of the invention may be engineered to includemodifications within the Fc region, typically to alter one or morefunctional properties of the antibody, such as serum half-life,complement fixation, Fc receptor binding, and/or antigen-dependentcellular cytotoxicity. Furthermore, an antibody of the invention may bechemically modified (e.g., one or more chemical moieties can be attachedto the antibody) or be modified to alter its glycosylation, again toalter one or more functional properties of the antibody. Each of theseembodiments is described in further detail below. The numbering ofresidues in the Fc region is that of the EU index of Kabat.

In one embodiment, the hinge region of CH1 is modified such that thenumber of cysteine residues in the hinge region is altered, e.g.,increased or decreased. This approach is described further in U.S. Pat.No. 5,677,425 by Bodmer et al. The number of cysteine residues in thehinge region of CH1 is altered to, for example, facilitate assembly ofthe light and heavy chains or to increase or decrease the stability ofthe antibody.

In another embodiment, the Fc hinge region of an antibody is mutated todecrease the biological half-life of the antibody. More specifically,one or more amino acid mutations are introduced into the CH2-CH3 domaininterface region of the Fc-hinge fragment such that the antibody hasimpaired Staphylococcyl protein A (SpA) binding relative to nativeFc-hinge domain SpA binding. This approach is described in furtherdetail in U.S. Pat. No. 6,165,745 by Ward et al.

In another embodiment, the antibody is modified to increase itsbiological half-life. Various approaches are possible. For example, oneor more of the following mutations can be introduced: T252L, T254S,T256F, as described in U.S. Pat. No. 6,277,375 to Ward. Alternatively,to increase the biological half life, the antibody can be altered withinthe CH1 or CL region to contain a salvage receptor binding epitope takenfrom two loops of a CH2 domain of an Fc region of an IgG, as describedin U.S. Pat. Nos. 5,869,046 and 6,121,022 by Presta et al.

In yet other embodiments, the Fc region is altered by replacing at leastone amino acid residue with a different amino acid residue to alter theeffector functions of the antibody. For example, one or more amino acidscan be replaced with a different amino acid residue such that theantibody has an altered affinity for an effector ligand but retains theantigen-binding ability of the parent antibody. The effector ligand towhich affinity is altered can be, for example, an Fc receptor or the C1component of complement. This approach is described in further detail inU.S. Pat. Nos. 5,624,821 and 5,648,260, both by Winter et al.

In another embodiment, one or more amino acids selected from amino acidresidues can be replaced with a different amino acid residue such thatthe antibody has altered C1q binding and/or reduced or abolishedcomplement dependent cytotoxicity (CDC). This approach is described infurther detail in U.S. Pat. No. 6,194,551 by Idusogie et al.

In another embodiment, one or more amino acid residues are altered tothereby alter the ability of the antibody to fix complement. Thisapproach is described further in PCT Publication WO 94/29351 by Bodmeret al.

In yet another embodiment, the Fc region is modified to increase theability of the antibody to mediate antibody dependent cellularcytotoxicity (ADCC) and/or to increase the affinity of the antibody foran Fcγ receptor by modifying one or more amino acids. This approach isdescribed further in PCT Publication WO 00/42072 by Presta. Moreover,the binding sites on human IgG1 for FcγRI, FcγRII, FcγRIII and FcRn havebeen mapped and variants with improved binding have been described (seeShields, R. L. et al., 2001 J. Biol. Chen. 276:6591-6604).

In still another embodiment, the glycosylation of an antibody ismodified. For example, an aglycoslated antibody can be made (i.e., theantibody lacks glycosylation). Glycosylation can be altered to, forexample, increase the affinity of the antibody for “antigen’. Suchcarbohydrate modifications can be accomplished by, for example, alteringone or more sites of glycosylation within the antibody sequence. Forexample, one or more amino acid substitutions can be made that result inelimination of one or more variable region framework glycosylation sitesto thereby eliminate glycosylation at that site. Such aglycosylation mayincrease the affinity of the antibody for antigen. Such an approach isdescribed in further detail in U.S. Pat. Nos. 5,714,350 and 6,350,861 byCo et al.

Additionally or alternatively, an antibody can be made that has analtered type of glycosylation, such as a hypofucosylated antibody havingreduced amounts of fucosyl residues or an antibody having increasedbisecting GlcNac structures. Such altered glycosylation patterns havebeen demonstrated to increase the ADCC ability of antibodies. Suchcarbohydrate modifications can be accomplished by, for example,expressing the antibody in a host cell with altered glycosylationmachinery. Cells with altered glycosylation machinery have beendescribed in the art and can be used as host cells in which to expressrecombinant antibodies of the invention to thereby produce an antibodywith altered glycosylation. For example, EP 1,176,195 by Hang et al.describes a cell line with a functionally disrupted FUT8 gene, whichencodes a fucosyl transferase, such that antibodies expressed in such acell line exhibit hypofucosylation. PCT Publication WO 03/035835 byPresta describes a variant CHO cell line, LecI3 cells, with reducedability to attach fucose to Asn(297)-linked carbohydrates, alsoresulting in hypofucosylation of antibodies expressed in that host cell(see also Shields, R. L. et al., 2002 J. Biol. Chem. 277:26733-26740).PCT Publication WO 99/54342 by Umana et al. describes cell linesengineered to express glycoprotein-modifying glycosyl transferases(e.g., beta(1,4)—N acetylglucosaminyltransferase III (GnTIII)) such thatantibodies expressed in the engineered cell lines exhibit increasedbisecting GlcNac structures which results in increased ADCC activity ofthe antibodies (see also Umana et al., 1999 Nat. Biotech. 17:176-180).

5.2.4. Methods of Engineering Altered Antibodies

As discussed above, the C5-binding antibodies having VH and VL sequencesor full length heavy and light chain sequences shown herein can be usedto create new C5-binding antibodies by modifying full length heavy chainand/or light chain sequences, VH and/or VL sequences, or the constantregion(s) attached thereto. Thus, in another aspect of the invention,the structural features of a C5-binding antibody of the invention areused to create structurally related C5-binding antibodies that retain atleast one functional property of the antibodies of the invention, suchas binding to human C5 and also inhibiting one or more functionalproperties of C5 (e.g., inhibit red blood cell lysis in a hemolyticassay).

For example, one or more CDR regions of the antibodies of the presentinvention, or mutations thereof, can be combined recombinantly withknown framework regions and/or other CDRs to create additional,recombinantly-engineered, C5-binding antibodies of the invention, asdiscussed above. Other types of modifications include those described inthe previous section. The starting material for the engineering methodis one or more of the VH and/or VL sequences provided herein, or one ormore CDR regions thereof. To create the engineered antibody, it is notnecessary to actually prepare (i.e., express as a protein) an antibodyhaving one or more of the VH and/or VL sequences provided herein, or oneor more CDR regions thereof. Rather, the information contained in thesequence(s) is used as the starting material to create a “secondgeneration” sequence(s) derived from the original sequence(s) and thenthe “second generation” sequence(s) is prepared and expressed as aprotein.

Accordingly, in another embodiment, the invention provides a method forpreparing an C5-binding antibody consisting of: a heavy chain variableregion antibody sequence having a CDR1 sequence selected from the groupconsisting of SEQ ID NOs: 1, 17, 33, 61, 131, 145, 159, 173, 195, and235, a CDR2 sequence selected from the group consisting of SEQ ID NOs:2, 18, 34, 49, 62, 77, 95, 107, 113, 119, 132, 146, 160, 174, 196, 226,and 236, and/or a CDR3 sequence selected from the group consisting ofSEQ ID NOs: 3, 19, 35, 63, 133, 147, 161, 175, 197, and 237; and a lightchain variable region antibody sequence having a CDR1 sequence selectedfrom the group consisting of SEQ ID NOs: 4, 20, 36, 64, 134, 148, 162,176, 198, and 238, a CDR2 sequence selected from the group consisting ofSEQ ID NOs: 5, 21, 37, 65, 135, 149, 163, 177, 199, and 239, and/or aCDR3 sequence selected from the group consisting of SEQ ID NOs: 6, 22,38, 50, 66, 78, 89, 101, 120, 136, 150, 164, 178, 200, 209, and 240;altering at least one amino acid residue within the heavy chain variableregion antibody sequence and/or the light chain variable region antibodysequence to create at least one altered antibody sequence; andexpressing the altered antibody sequence as a protein.

Accordingly, in another embodiment, the invention provides a method forpreparing an C5-binding antibody optimized for expression in a mammaliancell consisting of: a full length heavy chain antibody sequence having asequence selected from the group of SEQ ID NOs: 9, 25, 41, 53, 69, 81,97, 109, 115, 123, 139, 153, 167, 181, 189, 203, 212, 220, 229, 243,249, 254, 258, 274, 278, and 282; and a full length light chain antibodysequence having a sequence selected from the group of 10, 26, 42, 54,70, 82, 91, 103, 124, 140, 154, 168, 182, 190, 204, 213, 221, 230, 244,251, 262, 266, 270, 286, and 290; altering at least one amino acidresidue within the full length heavy chain antibody sequence and/or thefull length light chain antibody sequence to create at least one alteredantibody sequence; and expressing the altered antibody sequence as aprotein.

The altered antibody sequence can also be prepared by screening antibodylibraries having fixed CDR3 sequences or minimal essential bindingdeterminants as described in US20050255552 and diversity on CDR1 andCDR2 sequences. The screening can be performed according to anyscreening technology appropriate for screening antibodies from antibodylibraries, such as phage display technology.

Standard molecular biology techniques can be used to prepare and expressthe altered antibody sequence. The antibody encoded by the alteredantibody sequence(s) is one that retains one, some or all of thefunctional properties of the C5-binding antibodies described herein,which functional properties include, but are not limited to,specifically binding to human and/or cynomolgus C5; and the antibodyinhibit red blood cell lysis in a hemolytic assay.

The functional properties of the altered antibodies can be assessedusing standard assays available in the art and/or described herein, suchas those set forth in the Examples (e.g., ELISAs).

In certain embodiments of the methods of engineering antibodies of theinvention, mutations can be introduced randomly or selectively along allor part of an C5-binding antibody coding sequence and the resultingmodified C5-binding antibodies can be screened for binding activityand/or other functional properties as described herein. Mutationalmethods have been described in the art. For example, PCT Publication WO02/092780 by Short describes methods for creating and screening antibodymutations using saturation mutagenesis, synthetic ligation assembly, ora combination thereof. Alternatively, PCT Publication WO 03/074679 byLazar et al. describes methods of using computational screening methodsto optimize physiochemical properties of antibodies.

5.3. CHARACTERIZATION OF THE ANTIBODIES OF THE INVENTION

The antibodies of the invention can be characterized by variousfunctional assays. For example, they can be characterized by theirability to inhibit red blood cell lysis in hemolytic assays, theiraffinity to a C5 protein (e.g., human and/or cynomolgus C5), the epitopebinning, their resistance to proteolysis, and their ability to block thecomplement cascade, for example, their ability to inhibit MAC formation.

Various methods can be used to measure presence of complement pathwaymolecules and activation of the complement system (see, e.g., U.S. Pat.No. 6,087,120; and Newell et al., J Lab Clin Med, 100:437-44, 1982). Forexample, the complement activity can be monitored by (i) measurement ofinhibition of complement-mediated lysis of red blood cells (hemolysis);(ii) measurement of ability to inhibit cleavage of C3 or C5; and (iii)inhibition of alternative pathway mediated hemolysis.

The two most commonly used techniques are hemolytic assays (see, e.g.,Baatrup et al., Ann Rheum Dis, 51:892-7, 1992) and immunological assays(see, e.g., Auda et al., Rheumatol Int, 10:185-9, 1990). The hemolytictechniques measure the functional capacity of the entire sequence—eitherthe classical or alternative pathway. Immunological techniques measurethe protein concentration of a specific complement component or splitproduct. Other assays that can be employed to detect complementactivation or measure activities of complement components in the methodsof the present invention include, e.g., T cell proliferation assay(Chain et al., J Immunol Methods, 99:221-8, 1987), and delayed typehypersensitivity (DTH) assay (Forstrom et al., 1983, Nature 303:627-629;Halliday et al., 1982, in Assessment of Immune Status by the LeukocyteAdherence Inhibition Test, Academic, New York pp. 1-26; Koppi et al.,1982, Cell. Immunol. 66:394-406; and U.S. Pat. No. 5,843,449).

In hemolytic techniques, all of the complement components must bepresent and functional. Therefore hemolytic techniques can screen bothfunctional integrity and deficiencies of the complement system (see,e.g., Dijk et al., J Immunol Methods 36: 29-39, 1980; Minh et al., ClinLab Haematol. 5:23-34 1983; and Tanaka et al., J Immunol 86:161-170,1986). To measure the functional capacity of the classical pathway,sheep red blood cells coated with hemolysin (rabbit IgG to sheep redblood cells) or chicken red blood cells that are sensitized with rabbitanti-chicken antibodies are used as target cells (sensitized cells).These Ag-Ab complexes activate the classical pathway and result in lysisof the target cells when the components are functional and present inadequate concentration. To determine the functional capacity of thealternative pathway, rabbit red blood cells are used as the target cell(see, e.g., U.S. Pat. No. 6,087,120).

To test the ability of an antibody to inhibit MAC (membrance attackcomplex) formation, a MAC deposition assay can be performed. Briefly,zymosan can be used to activate the alternative pathway and IgM can beused to active the classic pathway. Fabs are pre-incubated with humanserum and added to plates coated with zymosan or IgM. Percentageinhibition of MAC deposition can be calculated for each sample relativeto baseline (EDTA treated human serum) and positive control (humanserum).

The ability of an antibody to bind to C5 can be detected by labellingthe antibody of interest directly, or the antibody may be unlabelled andbinding detected indirectly using various sandwich assay formats knownin the art.

In some embodiments, the C5-binding antibodies of the invention block orcompete with binding of a reference C5-binding antibody to a C5polypeptide. These can be fully human C5-binding antibodies describedabove. They can also be other mouse, chimeric or humanized C5-bindingantibodies which bind to the same epitope as the reference antibody. Thecapacity to block or compete with the reference antibody bindingindicates that a C5-binding antibody under test binds to the same orsimilar epitope as that defined by the reference antibody, or to anepitope which is sufficiently proximal to the epitope bound by thereference C5-binding antibody. Such antibodies are especially likely toshare the advantageous properties identified for the reference antibody.The capacity to block or compete with the reference antibody may bedetermined by, e.g., a competition binding assay. With a competitionbinding assay, the antibody under test is examined for ability toinhibit specific binding of the reference antibody to a common antigen,such as a C5 polypeptide. A test antibody competes with the referenceantibody for specific binding to the antigen if an excess of the testantibody substantially inhibits binding of the reference antibody.Substantial inhibition means that the test antibody reduces specificbinding of the reference antibody usually by at least 10%, 25%, 50%,75%, or 90%.

There are a number of known competition binding assays that can be usedto assess competition of a C5-binding antibody with the referenceC5-binding antibody for binding to a C5 protein. These include, e.g.,solid phase direct or indirect radioimmunoassay (RIA), solid phasedirect or indirect enzyme immunoassay (EIA), sandwich competition assay(see Stahli et al., Methods in Enzymology 9:242-253, 1983); solid phasedirect biotin-avidin EIA (see Kirkland et al., J. Immunol.137:3614-3619, 1986); solid phase direct labeled assay, solid phasedirect labeled sandwich assay (see Harlow & Lane, supra); solid phasedirect label RIA using I-125 label (see Morel et al., Molec. Immunol.25:7-15, 1988); solid phase direct biotin-avidin EIA (Cheung et al.,Virology 176:546-552, 1990); and direct labeled RIA (Moldenhauer et al.,Scand. J. Immunol. 32:77-82, 1990). Typically, such an assay involvesthe use of purified antigen bound to a solid surface or cells bearingeither of these, an unlabelled test C5-binding antibody and a labelledreference antibody. Competitive inhibition is measured by determiningthe amount of label bound to the solid surface or cells in the presenceof the test antibody. Usually the test antibody is present in excess.Antibodies identified by competition assay (competing antibodies)include antibodies binding to the same epitope as the reference antibodyand antibodies binding to an adjacent epitope sufficiently proximal tothe epitope bound by the reference antibody for steric hindrance tooccur.

To determine if the selected C5-binding monoclonal antibodies bind tounique epitopes, each antibody can be biotinylated using commerciallyavailable reagents (e.g., reagents from Pierce, Rockford, Ill.).Competition studies using unlabeled monoclonal antibodies andbiotinylated monoclonal antibodies can be performed using a C5polypeptide coated-ELISA plates. Biotinylated MAb binding can bedetected with a strep-avidin-alkaline phosphatase probe. To determinethe isotype of a purified C5-binding antibody, isotype ELISAs can beperformed. For example, wells of microtiter plates can be coated with 1μg/ml of anti-human IgG overnight at 4° C. After blocking with 1% BSA,the plates are reacted with 1 μg/ml or less of the monoclonal C5-bindingantibody or purified isotype controls, at ambient temperature for one totwo hours. The wells can then be reacted with either human IgG1 or humanIgM-specific alkaline phosphatase-conjugated probes. Plates are thendeveloped and analyzed so that the isotype of the purified antibody canbe determined.

To demonstrate binding of monoclonal C5-binding antibodies to live cellsexpressing a C5 polypeptide, flow cytometry can be used. Briefly, celllines expressing C5 (grown under standard growth conditions) can bemixed with various concentrations of a C5-binding antibody in PBScontaining 0.1% BSA and 10% fetal calf serum, and incubated at 37° C.for 1 hour. After washing, the cells are reacted withFluorescein-labeled anti-human IgG antibody under the same conditions asthe primary antibody staining. The samples can be analyzed by FACScaninstrument using light and side scatter properties to gate on singlecells. An alternative assay using fluorescence microscopy may be used(in addition to or instead of) the flow cytometry assay. Cells can bestained exactly as described above and examined by fluorescencemicroscopy. This method allows visualization of individual cells, butmay have diminished sensitivity depending on the density of the antigen.

C5-binding antibodies of the invention can be further tested forreactivity with a C5 polypeptide or antigenic fragment by Westernblotting. Briefly, purified C5 polypeptides or fusion proteins, or cellextracts from cells expressing C5 can be prepared and subjected tosodium dodecyl sulfate polyacrylamide gel electrophoresis. Afterelectrophoresis, the separated antigens are transferred tonitrocellulose membranes, blocked with 10% fetal calf serum, and probedwith the monoclonal antibodies to be tested. Human IgG binding can bedetected using anti-human IgG alkaline phosphatase and developed withBCIP/NBT substrate tablets (Sigma Chem. Co., St. Louis, Mo.).

Examples of functional assays are also described in the Example sectionbelow.

5.4. PROPHYLACTIC AND THERAPEUTIC USES

The present invention provides methods of treating a disease or disorderassociated with increased complement activity by administering to asubject in need thereof an effective amount of the antibodies of theinvention. In a specific embodiment, the present invention provides amethod of treating age-related macular degeneration (AMD) byadministering to a subject in need thereof an effective amount of theantibodies of the invention.

The antibodies of the invention can be used, inter alia, to preventprogression of dry AMD to wet AMD, to slow and/or prevent progression ofgeographic atrophy, and to improve vision lost due to dry AMDprogression. It can also be used in combination with anti-VEGF therapiesfor the treatment of wet AMD patients.

In some embodiments, the present invention provides methods of treatinga complement related disease or disorder by administering to a subjectin need thereof an effective amount of the antibodies of the invention.Examples of known complement related diseases or disorders include:neurological disorders, multiple sclerosis, stroke, Guillain BarreSyndrome, traumatic brain injury, Parkinson's disease, disorders ofinappropriate or undesirable complement activation, hemodialysiscomplications, hyperacute allograft rejection, xenograft rejection,interleukin-2 induced toxicity during IL-2 therapy, inflammatorydisorders, inflammation of autoimmune diseases, Crohn's disease, adultrespiratory distress syndrome, thermal injury including burns orfrostbite, post-ischemic reperfusion conditions, myocardial infarction,balloon angioplasty, post-pump syndrome in cardiopulmonary bypass orrenal bypass, hemodialysis, renal ischemia, mesenteric arteryreperfusion after acrotic reconstruction, infectious disease or sepsis,immune complex disorders and autoimmune diseases, rheumatoid arthritis,systemic lupus erythematosus (SLE), SLE nephritis, proliferativenephritis, hemolytic anemia, and myasthenia gravis. In addition, otherknown complement related disease are lung disease and disorders such asdyspnea, hemoptysis, ARDS, asthma, chronic obstructive pulmonary disease(COPD), emphysema, pulmonary embolisms and infarcts, pneumonia,fibrogenic dust diseases, inert dusts and minerals (e.g., silicon, coaldust, beryllium, and asbestos), pulmonary fibrosis, organic dustdiseases, chemical injury (due to irritant gasses and chemicals, e.g.,chlorine, phosgene, sulfur dioxide, hydrogen sulfide, nitrogen dioxide,ammonia, and hydrochloric acid), smoke injury, thermal injury (e.g.,burn, freeze), asthma, allergy, bronchoconstriction, hypersensitivitypneumonitis, parasitic diseases, Goodpasture's Syndrome, pulmonaryvasculitis, and immune complex-associated inflammation.

In a specific embodiment, the present invention provides methods oftreating a complement related disease or disorder by administering to asubject in need thereof an effective amount of the antibodies of theinvention, wherein said disease or disorder is asthma, arthritis (e.g.,rheumatoid arthritis), autoimmune heart disease, multiple sclerosis,inflammatory bowel disease, ischemia-reperfusion injuries,Barraquer-Simons Syndrome, hemodialysis, systemic lupus, lupuserythematosus, psoriasis, multiple sclerosis, transplantation, diseasesof the central nervous system such as Alzheimers disease and otherneurodegenerative conditions, aHUS, glomerulonephritis, bullouspemphigoid or MPGN II.

In a specific embodiment, the present invention provides methods oftreating glomerulonephritis by administering to a subject in needthereof an effective amount of a composition comprising an antibody ofthe present invention. Symptoms of glomerulonephritis include, but notlimited to, proteinuria; reduced glomerular filtration rate (GFR); serumelectrolyte changes including azotemia (uremia, excessive blood ureanitrogen—BUN) and salt retention, leading to water retention resultingin hypertension and edema; hematuria and abnormal urinary sedimentsincluding red cell casts; hypoalbuminemia; hyperlipidemia; andlipiduria. In a specific embodiment, the present invention providesmethods of treating paroxysmal nocturnal hemoglobinuria (PNH byadministering to a subject in need thereof an effective amount of acomposition comprising an antibody of the present invention.

In a specific embodiment, the present invention provides methods ofreducing the dysfunction of the immune and hemostatic systems associatedwith extracorporeal circulation by administering to a subject in needthereof an effective amount of a composition comprising an antibody ofthe present invention. The antibodies of the present invention can beused in any procedure which involves circulating the patient's bloodfrom a blood vessel of the patient, through a conduit, and back to ablood vessel of the patient, the conduit having a luminal surfacecomprising a material capable of causing at least one of complementactivation, platelet activation, leukocyte activation, orplatelet-leukocyte adhesion. Such procedures include, but are notlimited to, all forms of ECC, as well as procedures involving theintroduction of an artificial or foreign organ, tissue, or vessel intothe blood circuit of a patient.

Subjects to be treated with therapeutic agents of the present inventioncan also be administered other therapeutic agents with know methods oftreating conditions associated with macular degeneration, such asantibiotic treatments as described in U.S. Pat. No. 6,218,368. In othertreatments, immunosuppressive agents such as cyclosporine, are agentscapable of suppressing immune responses. These agents include cytotoxicdrugs, corticosteroids, nonsteroidal anti-inflammatory drugs (NSAIDs),specific T-lymphocyte immunosuppressants, and antibodies or fragmentsthereof (see Physicians' Desk Reference, 53rd edition, Medical EconomicsCompany Inc., Montvale, N.J. (1999). Immunosuppressive treatment istypically continued at intervals for a period of a week, a month, threemonths, six months or a year. In some patients, treatment isadministered for up to the rest of a patient's life.

When the therapeutic agents of the present invention are administeredtogether with another agent, the two can be administered sequentially ineither order or simultaneously. In some aspects, an antibody of thepresent invention is administered to a subject who is also receivingtherapy with a second agent (e.g., verteporfin). In other aspects, thebinding molecule is administered in conjunction with surgicaltreatments.

Suitable agents for combination treatment with C5-binding antibodiesinclude agents known in the art that are able to modulate the activitiesof complement components (see, e.g., U.S. Pat. No. 5,808,109). Otheragents have been reported to diminish complement-mediated activity. Suchagents include: amino acids (Takada, Y. et al. Immunology 1978, 34,509); phosphonate esters (Becker, L. Biochem. Biophy. Acta 1967, 147,289); polyanionic substances (Conrow, R. B. et al. J. Med. Chem. 1980,23, 242); sulfonyl fluorides (Hansch, C.; Yoshimoto, M. J. Med. Chem.1974, 17, 1160, and references cited therein); polynucleotides(DeClercq, P. F. et al. Biochem. Biophys. Res. Commun. 1975, 67, 255);pimaric acids (Glovsky, M. M. et al. J. Immunol. 1969, 102, 1);porphines (Lapidus, M. and Tomasco, J. Immunopharmacol. 1981, 3, 137);several antiinflammatories (Burge, J. J. et a! J. Immunol. 1978, 120,1625); phenols (Muller-Eberhard, H. J. 1978, in Molecular Basis ofBiological Degradative Processes, Berlin, R. D. et al., eds. AcademicPress, New York, p. 65); and benzamidines (Vogt, W. et al Immunology1979, 36, 138). Some of these agents function by general inhibition ofproteases and esterases. Others are not specific to any particularintermediate step in the complement pathway, but, rather, inhibit morethan one step of complement activation. Examples of the latter compoundsinclude the benzamidines, which block C1, C4 and C5 utilization (see,e.g., Vogt et al. Immunol. 1979, 36, 138).

Additional agents known in the art that can inhibit activity ofcomplement components include K-76, a fungal metabolite fromStachybotrys (Corey et al., J. Amer. Chem. Soc. 104: 5551, 1982). BothK-76 and K-76 COOH have been shown to inhibit complement mainly at theC5 step (Hong et al., J. Immunol. 122: 2418, 1979; Miyazaki et al.,Microbiol. Immunol. 24: 1091, 1980), and to prevent the generation of achemotactic factor from normal human complement (Bumpers et al., Lab.Clinc. Med. 102: 421, 1983). At high concentrations of K-76 or K-76COOH, some inhibition of the reactions of C2, C3, C6, C7, and C9 withtheir respective preceding intermediaries is exhibited. K-76 or K-76COOH has also been reported to inhibit the C3b inactivator system ofcomplement (Hong et al., J. Immunol. 127: 104-108, 1981). Other suitableagents for practicing methods of the present invention includegriseofulvin (Weinberg, in Principles of Medicinal Chemistry, 2d Ed.,Foye, W. O., ed., Lea & Febiger, Philadelphia, Pa., p. 813, 1981),isopannarin (Djura et al., Aust. J. Chem. 36: 1057, 1983), andmetabolites of Siphonodictyon coralli-phagum (Sullivan et al.,Tetrahedron 37: 979, 1981).

A combination therapy regimen may be additive, or it may producesynergistic results (e.g., reductions in complement pathway activitymore than expected for the combined use of the two agents). In someembodiments, the present invention provide a combination therapy forpreventing and/or treating AMD or another complement related disease asdescribed above with a C5-binding antibody of the invention and ananti-angiogenic, such as anti-VEGF agent.

5.5. DIAGNOSTIC USES

In one aspect, the invention encompasses diagnostic assays fordetermining C5 protein and/or nucleic acid expression as well as C5protein function, in the context of a biological sample (e.g., blood,serum, cells, tissue) or from individual is afflicted with a disease ordisorder, or is at risk of developing a disorder associated with AMD.

Diagnostic assays, such as competitive assays rely on the ability of alabelled analogue (the “tracer”) to compete with the test sample analytefor a limited number of binding sites on a common binding partner. Thebinding partner generally is insolubilized before or after thecompetition and then the tracer and analyte bound to the binding partnerare separated from the unbound tracer and analyte. This separation isaccomplished by decanting (where the binding partner waspreinsolubilized) or by centrifuging (where the binding partner wasprecipitated after the competitive reaction). The amount of test sampleanalyte is inversely proportional to the amount of bound tracer asmeasured by the amount of marker substance. Dose-response curves withknown amounts of analyte are prepared and compared with the test resultsin order to quantitatively determine the amount of analyte present inthe test sample. These assays are called ELISA systems when enzymes areused as the detectable markers. In an assay of this form, competitivebinding between antibodies and C5-binding antibodies results in thebound C5 protein, preferably the C5 epitopes of the invention, being ameasure of antibodies in the serum sample, most particularly,neutralising antibodies in the serum sample.

A significant advantage of the assay is that measurement is made ofneutralising antibodies directly (i.e., those which interfere withbinding of C5 protein, specifically, epitopes). Such an assay,particularly in the form of an ELISA test has considerable applicationsin the clinical environment and in routine blood screening.

Immunologic techniques employ polyclonal or monoclonal antibodiesagainst the different epitopes of the various complement components(e.g., C3, C4, C5) to detect, e.g., the split products of complementcomponents (see, e.g., Hugli et al., Immunoassays Clinical LaboratoryTechniques 443-460, 1980; Gorski et al., J Immunol Meth 47: 61-73, 1981;Linder et al., J Immunol Meth 47: 49-59, 1981; and Burger et al., JImmunol 141: 553-558, 1988). Binding of the antibody with the splitproduct in competition with a known concentration of labeled splitproduct could then be measured. Various assays such asradio-immunoassays, ELISA's, and radial diffusion assays are availableto detect complement split products.

The immunologic techniques provide high sensitivity to detect complementactivation, since they allow measurement of split-product formation inblood from a test subject and control subjects with or without maculardegeneration-related disorders. Accordingly, in some methods of thepresent invention, diagnosis of a disorder associated with oculardisorders is obtained by measurement of abnormal complement activationthrough quantification of the soluble split products of complementcomponents in blood plasma from a test subject. The measurements can beperformed as described, e.g., in Chenoweth et al., N Engl J Med 304:497-502, 1981; and Bhakdi et al., Biochim Biophys Acta 737: 343-372,1983. Preferably, only the complement activation formed in vivo ismeasured. This can be accomplished by collecting a biological samplefrom the subject (e.g., serum) in medium containing inhibitors of thecomplement system, and subsequently measuring complement activation(e.g., quantification of the split products) in the sample.

In the clinical diagnosis or monitoring of patients with disordersassociated with ocular diseases or disorders, the detection ofcomplement proteins in comparison to the levels in a correspondingbiological sample from a normal subject is indicative of a patient withdisorders associated with macular degeneration.

In vivo diagnostic or imaging is described in US2006/0067935. Briefly,these methods generally comprise administering or introducing to apatient a diagnostically effective amount of a C5 binding molecule thatis operatively attached to a marker or label that is detectable bynon-invasive methods. The antibody-marker conjugate is allowedsufficient time to localize and bind to complement proteins within theeye. The patient is then exposed to a detection device to identify thedetectable marker, thus forming an image of the location of the C5binding molecules in the eye of a patient. The presence of C5 bindingantibody or an antigen-binding fragment thereof is detected bydetermining whether an antibody-marker binds to a component of the eye.Detection of an increased level in selected complement proteins or acombination of protein in comparison to a normal individual without AMDdisease is indicative of a predisposition for and/or on set of disordersassociated with macular degeneration. These aspects of the invention arealso preferred for use in eye imaging methods and combined angiogenicdiagnostic and treatment methods.

The invention also pertains to the field of predictive medicine in whichdiagnostic assays, prognostic assays, pharmacogenomics, and monitoringclinical trials are used for prognostic (predictive) purposes to therebytreat an individual prophylactically.

The invention also provides for prognostic (or predictive) assays fordetermining whether an individual is at risk of developing a disorderassociated with dysregulation of complement pathway activity. Forexample, mutations in a C5 gene can be assayed in a biological sample.Such assays can be used for prognostic or predictive purpose to therebyprophylactically treat an individual prior to the onset of a disordercharacterized by or associated with C5 protein, nucleic acid expressionor activity.

Another aspect of the invention provides methods for determining C5nucleic acid expression or C5 protein activity in an individual tothereby select appropriate therapeutic or prophylactic agents for thatindividual (referred to herein as “pharmacogenomics”). Pharmacogenomicsallows for the selection of agents (e.g., drugs) for therapeutic orprophylactic treatment of an individual based on the genotype of theindividual (e.g., the genotype of the individual examined to determinethe ability of the individual to respond to a particular agent.)

Yet another aspect of the invention pertains to monitoring the influenceof agents (e.g., drugs) on the expression or activity of C5 protein inclinical trials.

5.6. PHARMACEUTICAL COMPOSITIONS

The invention provides pharmaceutical compositions comprising theC5-binding antibodies (intact or binding fragments) formulated togetherwith a pharmaceutically acceptable carrier. The compositions canadditionally contain one or more other therapeutical agents that aresuitable for treating or preventing a complement-associated disease(e.g., AMD). Pharmaceutically carriers enhance or stabilize thecomposition, or to facilitate preparation of the composition.Pharmaceutically acceptable carriers include solvents, dispersion media,coatings, antibacterial and antifungal agents, isotonic and absorptiondelaying agents, and the like that are physiologically compatible.

A pharmaceutical composition of the present invention can beadministered by a variety of methods known in the art. The route and/ormode of administration vary depending upon the desired results. It ispreferred that administration be intravenous, intramuscular,intraperitoneal, or subcutaneous, or administered proximal to the siteof the target. In a specific embodiment, the antibodies of the inventionare formulated so that they can be administered intravitreally into theeye. The pharmaceutically acceptable carrier should be suitable forintravenous, intramuscular, subcutaneous, parenteral, spinal orepidermal administration (e.g., by injection or infusion). Depending onthe route of administration, the active compound, i.e., antibody,bispecific and multispecific molecule, may be coated in a material toprotect the compound from the action of acids and other naturalconditions that may inactivate the compound.

The composition should be sterile and fluid. Proper fluidity can bemaintained, for example, by use of coating such as lecithin, bymaintenance of required particle size in the case of dispersion and byuse of surfactants. In many cases, it is preferable to include isotonicagents, for example, sugars, polyalcohols such as mannitol or sorbitol,and sodium chloride in the composition. Long-term absorption of theinjectable compositions can be brought about by including in thecomposition an agent which delays absorption, for example, aluminummonostearate or gelatin.

Pharmaceutical compositions of the invention can be prepared inaccordance with methods well known and routinely practiced in the art.See, e.g., Remington: The Science and Practice of Pharmacy, MackPublishing Co., 20th ed., 2000; and Sustained and Controlled ReleaseDrug Delivery Systems, J. R. Robinson, ed., Marcel Dekker, Inc., NewYork, 1978. Pharmaceutical compositions are preferably manufacturedunder GMP conditions. Typically, a therapeutically effective dose orefficacious dose of the C5-binding antibody is employed in thepharmaceutical compositions of the invention. The C5-binding antibodiesare formulated into pharmaceutically acceptable dosage forms byconventional methods known to those of skill in the art. Dosage regimensare adjusted to provide the optimum desired response (e.g., atherapeutic response). For example, a single bolus may be administered,several divided doses may be administered over time or the dose may beproportionally reduced or increased as indicated by the exigencies ofthe therapeutic situation. It is especially advantageous to formulateparenteral compositions in dosage unit form for ease of administrationand uniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the subjects tobe treated; each unit contains a predetermined quantity of activecompound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier.

Actual dosage levels of the active ingredients in the pharmaceuticalcompositions of the present invention can be varied so as to obtain anamount of the active ingredient which is effective to achieve thedesired therapeutic response for a particular patient, composition, andmode of administration, without being toxic to the patient. The selecteddosage level depends upon a variety of pharmacokinetic factors includingthe activity of the particular compositions of the present inventionemployed, or the ester, salt or amide thereof, the route ofadministration, the time of administration, the rate of excretion of theparticular compound being employed, the duration of the treatment, otherdrugs, compounds and/or materials used in combination with theparticular compositions employed, the age, sex, weight, condition,general health and prior medical history of the patient being treated,and like factors.

A physician or veterinarian can start doses of the antibodies of theinvention employed in the pharmaceutical composition at levels lowerthan that required to achieve the desired therapeutic effect andgradually increase the dosage until the desired effect is achieved. Ingeneral, effective doses of the compositions of the present invention,for the treatment of an allergic inflammatory disorder described hereinvary depending upon many different factors, including means ofadministration, target site, physiological state of the patient, whetherthe patient is human or an animal, other medications administered, andwhether treatment is prophylactic or therapeutic. Treatment dosages needto be titrated to optimize safety and efficacy. For systemicadministration with an antibody, the dosage ranges from about 0.0001 to100 mg/kg, and more usually 0.01 to 15 mg/kg, of the host body weight.An exemplary treatment regime entails systemic administration once perevery two weeks or once a month or once every 3 to 6 months. Forintravitreal administration with an antibody, the dosage ranges fromabout 0.0001 to about 10 mg. An exemplary treatment regime entailssystemic administration once per every two weeks or once a month or onceevery 3 to 6 months.

Antibody is usually administered on multiple occasions. Intervalsbetween single dosages can be weekly, monthly or yearly. Intervals canalso be irregular as indicated by measuring blood levels of C5-bindingantibody in the patient. In some methods of systemic administration,dosage is adjusted to achieve a plasma antibody concentration of 1-1000μg/ml and in some methods 25-500 μg/ml. Alternatively, antibody can beadministered as a sustained release formulation, in which case lessfrequent administration is required. Dosage and frequency vary dependingon the half-life of the antibody in the patient. In general, humanizedantibodies show longer half life than that of chimeric antibodies andnonhuman antibodies. The dosage and frequency of administration can varydepending on whether the treatment is prophylactic or therapeutic. Inprophylactic applications, a relatively low dosage is administered atrelatively infrequent intervals over a long period of time. Somepatients continue to receive treatment for the rest of their lives. Intherapeutic applications, a relatively high dosage at relatively shortintervals is sometimes required until progression of the disease isreduced or terminated, and preferably until the patient shows partial orcomplete amelioration of symptoms of disease. Thereafter, the patientcan be administered a prophylactic regime.

6. EXAMPLES

The following examples are provided to further illustrate the inventionbut not to limit its scope. Other variants of the invention will bereadily apparent to one of ordinary skill in the art and are encompassedby the appended claims.

Example 1 Generation of Cynomolgus C5 and Human C5 1. Generation ofCynomolgus C5

Cynomolgus C5 was purified successfully from cynomolgus serum byaffinity chromatography using MOR07086 hu IgG1. Cynomolgus C5 wasquality tested by SDS-PAGE, Western blot, mass spectrometry andhemolytic assays. Quality of purified cynomolgus C5 was shown to be highby SDS-PAGE and Western blot. Lack of C3 contamination was confirmed bySDS and Western blot. In addition, the identity of cynomolgus C5sequence was determined by mass spectrometric analysis and the activityof purified cynomolgus C5 was tested in hemolytic assays. In hemolyticassays the new preparation was equipotent to human C5 (e.g., Sample 6,which was used in affinity maturation pannings, reconstituted complementactivity of 20% human C5-depleted serum with similar activity topurified human C5).

2. Quality Control of Human and Cynomolgus Biotinylated andNon-Biotinylated C5 Proteins

Bioactivity of purified human C5 was characterized and confirmed by thealternative pathway hemolytic activity. C5 was spiked into C5-depletedhuman serum at varying concentrations to obtain an EC50. EC50 valuesranging between 0.02-0.1 nM were considered acceptable.

Before using, the bioactivity of every purified human C5 protein lot wastested in the hemolytic assay. The same quality control was done forcynomolgus C5 after purification from cynomolgus serum. Afterbiotinylation of human and cynomolgus C5, the bioactivity of thematerial was also tested in hemolytic assays, in order to analyze ifthere was a loss of activity caused by biotinylation.

Example 2 Generation of C5-Specific Antibodies from the HuCAL GOLD®Library

C5 antibodies were generated by selection of clones having high bindingaffinities, using as the source of antibody variant proteins acommercially available phage display library, the MorphoSys HuCAL GOLD®library.

HuCAL GOLD® library is a Fab library (Knappik et al., 2000) in which allsix CDRs are diversified by appropriate mutation, and which employs theCysDisplay™ technology for linking the Fab to the phage surface(WO01/05950, Löhning et al., 2001).

1. Selection by Panning of C5 Specific Antibodies from the Library

For the selection of antibodies against C5, two different panningstrategies were applied. The six different pools were individuallysubjected to three rounds of: (a) a solid phase panning where theantigens (human and cynomolgus C5) were directly coated on Maxisorp 96well microtiter plates (Nunc, Wiesbaden, Germany); or (b) a solutionpanning with biotinylated human and cyno C5 where the phage-antigencomplex was captured by Streptavidin magnetic beads (Dynabeads M-280;Dynal) for each panning pool.

The HuCAL GOLD® library was amplified in 2×YT medium containing 34 μg/mlchloramphenicol and 1% glucose (2×YT-CG). After infection with VCSM13helper phage at an OD600 nm of 0.5 (30 min at 37° C. without shaking; 30min at 37° C. shaking at 250 rpm), cells were spun down (4120 g; 5 min;4° C.), resuspended in 2×YT/34 μg/ml chloramphenicol/50 μg/mlkanamycin/0.25 mM IPTG and grown overnight at 22° C. Phage werePEG-precipitated from the supernatant, resuspended in PBS/20% glyceroland stored at −80° C. Phage amplification between two panning rounds wasconducted as follows: mid-log phase E. coli TG1 cells were infected witheluted phage and plated onto LB-agar supplemented with 1% of glucose and34 μg/ml of chloramphenicol (LB-CG plates). After overnight incubationat 30° C., the TG1 colonies were scraped off the agar plates and used toinoculate 2×YT-CG until an OD600 nm of 0.5 was reached. VCSM13 helperphage were added for infection as described above.

Taken together 354 clones derived from all panning strategies weresequenced, resulting in 64 unique clones with the desired profile:binding to human and cynomolgus C5 and no binding to the counter targetsC3 and C4.

45 clones derived from solid phase pannings and 19 clones from solutionpannings were selected for protein expression and purification. FourFabs from solid phase pannings (MOR06525, MOR06756, MOR06757 andMOR06763) and 6 Fabs from solution pannings (MOR07086, MOR07087,MOR07091, MOR07092, MOR07093 and MOR07094) entered affinity maturation.

Solid Phase Panning Against C5 on Directly Coated Protein

The first panning variant was solid phase panning alternating human C5(first and third round of selection) and cynomolgus C5 (second round ofselection).

Three wells of a Maxisorp plate (F96 Nunc-Immunoplate) were coated with200 μl of 50 nM C5 each o/n at 4° C. The coated wells were washed 2×with 400 μl PBS and blocked with 350 μl 5% MPBS for 2 h at RT on amicrotiter plate shaker. For each panning about 10¹³ HuCAL GOLD®phage-antibodies were blocked with equal volume of PBST/5% milk powderfor 2 h at room temperature. The coated wells were washed 2× with 400 μlPBS after the blocking procedure. 200 μl of pre-blocked HuCAL GOLD®phage-antibodies were added to each coated well and incubated for 2 h atRT on a shaker. Washing was performed by adding five times 350 μlPBS/0.05% Tween, followed by washing another five times with PBS. Forsome panning conditions a more stringent wash procedure was applied.

Elution of phage from the plate was performed with 200 μl 20 mM DTT in10 mM Tris/HCl pH8 per well for 10 min. The DTT phage eluate was addedto 15 ml of E. coli TG1, which were grown to an OD600 of 0.6-0.8 at 37°C. in 2YT medium and incubated in 50 ml plastic tubes for 45 min at 37°C. without shaking for phage infection. After centrifugation for 5 minat 4120×g, the bacterial pellets were each resuspended in 600 μl 2×YTmedium, plated on 3×YT-CG agar plates and incubated overnight at 37° C.Colonies were scraped off the plates and phages were rescued andamplified as described above.

The second and third rounds of solid phase panning were performedaccording to the protocol of the first round. In the second selectionround for some panning conditions the output of the first round was usedfor selections on cynomolgus C5 in order to enrich for cynomolguscross-reactive antibodies.

For some panning conditions washing stringency was increased and antigenconcentration was decreased within the three round of selection in orderto generate high affinity antibodies.

The HuCAL GOLD® phagemid library was used to select specific Fabantibody fragments against human C5. First strategy was a solid phasepanning on directly coated human C5 protein (panning procedure describedabove).

After the 3rd panning round, the enriched phage pools were subclonedfrom the pMORPH®23 library vector (allowing efficient antibody displayon the phage surface) into the pMORPH®x9_Fab_MH expression vector whichmediates periplasmic expression of soluble Fabs. Single clones werepicked and soluble Fabs were expressed from these single clones.

In total, 6624 clones were analyzed in primary screening which wasperformed by binding of the Fabs directly from the bacterial lysates tohuman C5 immobilized on Maxisorp microtiter plates. 1660 hits wereobtained from the primary screening on human C5 with signals >5-foldover background. 384 hits were further analyzed in a secondary screeningto confirm binding on human C5 and to screen for binding to the countertargets human C3 and C4.

Many primary hits could be confirmed on human C5 and showed nocross-reactivity to human C3 and C4, but only 6 Fabs had weakcross-reactivity to cynomolgus C5.

As a first consequence new solid phase pannings were performedalternating on human and cynomolgus C5. In parallel, quality controls ofthe purified cynomolgus C5 batch revealed a high amount of cynomolgus C3within the cynomolgus C5 batch. Considering this results, a new methodto screen for cynomolgus cross-reactive antibodies was applied.Cynomolgus C5 was captured from cynomolgus serum using an C5-bindingpolyclonal antibody (see Example 3, section 3). Using this method theinitial primary hits were screened again on cynomolgus C5 and 56 cloneswere confirmed for binding to cynomolgus C5.

For the alternating solid phase pannings, the 1st round output of themost successful 12 human solid phase pannings was used for selections oncynomolgus C5 (protein batch contaminated with cynomolgus C3; not knownduring pannings). 376 clones were confirmed in a secondary screening forbinding to human C5 and 361 clones for binding to cynomolgus C5 capturedfrom cynomolgus serum.

Solution Panning on Biotinylated C5 Protein

The second panning variant was solution panning against biologicallyactive (in hemolytic assays) biotinylated human C5 and biotinylatedcynomolgus C5.

For this panning 200 μl of Streptavidin magnetic beads (Dynabeads M-280;Dynal) were washed once with PBS and blocked with Chemiblocker for 2 hat RT. 300 μl of the PBS diluted phage were blocked also withChemiblocker for 1-2 h at RT on a rotator. The blocked phages were twicepre-adsorbed against 50 μl blocked Streptavidin magnetic beads for 30min. The phage supernatant was transferred to a new blocked 2 mlreaction tube and human biotinylated C5 was added and incubated for 1 hat RT on a rotator. 100 μl of the blocked Streptavidin magnetic beadswere added to each panning pool an incubated for 10 min on a rotator.The beads were collected with a particle separator (Dynal MPC-E) forapprox. 2.5 min and the solution was removed carefully.

Beads were then washed 7× in PBS/0.05% Tween using a rotator, followedby washing another three times with PBS. Elution of phage from theDynabeads was performed by adding 200 μl 20 mM DTT in 10 mM Tris/HCl pH8 to each tube and incubation for 10 min. Dynabeads were removed by themagnetic particle separator and the supernatant was added to 15 ml of anE. coli TG-1 culture grown to OD600 nm of 0.6-0.8. Beads were thenwashed once with 200 μl PBS and together with additionally removedphages the PBS was added to the 15 ml E. coli TG-1 culture. For phageinfection, the culture was incubated in 50 ml plastic tubes for 45 minat 37° C. without shaking. After centrifugation for 5 min at 4120×g, thebacterial pellets were resuspended each in 600 μl 2×YT medium, plated on3×YT-CG agar plates and incubated overnight at 37° C. Colonies werescraped off the plates and phages were rescued and amplified asdescribed above. The second and third rounds of selection were performedin an identical way to the first round of selection.

A further panning strategy was solution panning using human C5 andalternating human and cynomolgus C5 (protein batch contaminated withcynomolgus C3, not known during pannings). Therefore the proteins werebiotinylated and the retained bio-functionality after the biotinylationprocedure was confirmed in hemolytic bioassays.

The phage-antigen complex was captured on Streptavidin magnetic beadsvia the biotin moiety of the antigen. After washing only specific boundphage were eluted (panning procedure described above).

First screening was done on directly coated proteins (see Example 3,section 1) and only 80 clones could be confirmed on human C5. Due to thefact that during the pannings the antigen was kept in solution, a newscreening method was developed. In a solution ELISA the Fabs wereincubated with biotinylated antigen on a NeutrAvidin plate. Using thissolution screening procedure, a significantly higher amount of human andcynomolgus C5 specific clones could be selected. These results confirmedthat many Fabs derived from solution pannings recognize C5 only insolution or when captured (e.g. via a polyclonal C5-binding antibody).

2. Subcloning and Expression of Selected Fab Fragments

To facilitate rapid expression of soluble Fabs, the Fab-encoding insertsof the selected HuCAL GOLD® phages were subcloned via XbaI and EcoRIinto the E. coli expression vector pMORPH®x9_MH. Fab fragments carry aC-terminal Myc tag and as a second C-terminal tag the 6×His-tag (Chen etal., Gene 139:73-75 (1994)). After transformation of the expressionplasmids into E. coli TG1 F− cells chloramphenicol-resistant singleclones were picked into the wells of a sterile 384-well microtiter platepre-filled with 60 μl 2×YT-CG medium and grown o/n at 30° C. 5 μl ofeach E. coli TG-1 o/n culture were transferred to a fresh, sterile96-well microtiter plate pre-filled with 40 μl 2×YT medium supplementedwith 34 μg/ml chloramphenicol per well. The microtiter plates wereincubated at 30° C. shaking at 400 rpm on a microplate shaker until thecultures were slightly turbid (˜2-4 h) with an OD600 nm of ˜0.5. Tothese expression plates, 10 μl 2×YT medium supplemented with 34 μg/mlchloramphenicol and 3 mM IPTG (isopropyl-β-D-thiogalactopyranoside) wasadded per well (end concentration 0.5 mM IPTG). The microtiter plateswere sealed with a gas-permeable tape, and incubated o/n at 30° C.shaking at 400 rpm. To each well of the expression plates, 15 μl BELbuffer was added containing 2.5 mg/ml lysozyme, 4 mM EDTA and 10 U/μlBenzonase and incubated for 1 h at 22° C. on a microtiter plate shaker(400 rpm) followed by an optional freezing step for at least 2 h at −80°C. The BEL extracts were used for binding analysis by ELISA or Fab SETscreening after affinity maturation.

Expression of Fab fragments encoded by pMORPH®x9_Fab_MH in TG-1 cellswas carried out in shaker flask cultures using 750 ml of 2×YT mediumsupplemented with 34 μg/ml chloramphenicol. Cultures were shaken at 30°C. until the OD600 nm reached 0.5. Expression was induced by addition of0.75 mM IPTG for 20 h at 30° C. Cells were disrupted using lysozyme andFab fragments isolated by Ni-NTA chromatography (Qiagen, Hilden,Germany). Buffer exchange to 1× Dulbecco's PBS (pH 7.2) was performedusing PD10 columns. Samples were filtered sterile (0.2 μm, Millipore).Purity of samples was determined in denatured, reduced state by SDS-PAGE(15% Criterion Gels, BioRad) and in native state by size exclusionchromatography (HP-SEC). Protein concentrations were determined byUV-spectrophotometry (Krebs et al., J. Immunol. Methods 254:67-84(2001)).

On Fab level, the overall expression rates and the percentage ofmonomeric fraction in SEC (Size Exclusion Chromatography) ranged fromacceptable to good for most of the identified antibody fragments. 64parental Fabs were expressed and 61 Fabs could be purified. 60 affinitymatured Fabs were purified in the mg scale. Most of the Fabs were goodexpressors and had no aggregation tendency.

Example 3 Identification of C5-Specific Antibodies from the HuCAL Gold®Library

Below four different Enzyme Linked Immunosorbent Assay (ELISA) methodsdescribe the screening of C5-binding antibodies (as bacterial BELlysates or purified Fabs) on specific and counter antigens.

1. Screening on Directly Coated Protein

Maxisorp (Nunc, Rochester, N.Y., USA) 384 well plates were coated with20 μl per well of 2.5 μg/ml antigen (human C5 and the counter proteinshuman C3 and C4) in PBS, pH 7.4 o/n at 4° C. In parallel, plates werecoated with 20 μl per well of 5 μg/ml sheep anti-human IgG, Fd fragmentspecific (The Binding Site, Birmingham, UK), diluted in PBS, pH 7.4 tocheck for Fab expression level.

The plates were blocked with PBS/0.05% Tween 20 (PBST) containing 5%milk powder for 1-2 h at RT. After washing the wells with PBST,BEL-extracts, purified HuCAL GOLD® Fabs or control Fabs diluted in PBSwere added and incubated for 1 h at RT. To detect Fab binding, anti-HIS6antibody coupled to peroxidase was applied (Roche).

For detection of POD-conjugates fluorogenic substrate QuantaBlu (Pierce)was used according to manufacturer's instructions. Between allincubation steps, the wells of the microtiter plates were washed threetimes and five times with PBST after the final incubation with thesecondary antibody. Fluorescence was measured in a Tecan GENios Proplate reader.

2. Solution Screening with Biotinylated Proteins

The ELISA method described below was used for screening of HuCAL GOLD®Fabs after solution pannings using biotinylated complement proteins.

NeutrAvidin plates were blocked with 1× Chemiblocker (Chemicon) dilutedin PBS o/n at 4° C. These plates were used to screen for binding tohuman C5 and to the counter targets C3 and C4. In parallel, Maxisorp 384well plates (Nunc, Rochester, N.Y., USA) were coated with 20 μl per wellof 5 μg/ml sheep anti-human IgG, Fd fragment specific (The Binding Site,Birmingham, UK), diluted in PBS, pH 7.4. These plates were used to checkfor Fab expression levels and for non-specific biotin binding. On thenext day, coated Maxisorp plates were washed 2× with PBST and blockedwith 3% BSA in TBS for 1-2 h at RT. Periplasmic BEL extracts containingFabs or purified HuCAL GOLD® Fabs were added to both blocked NeutrAvidinand Maxisorp plates.

Subsequently, 20 μl per well of biotinylated human C5 (to detectspecific binding) and in parallel, biotinylated human C3 and C4 (todetect unwanted binding) were added to wells of the NeutrAvidin plates.The biotinylated antigens were incubated with the HuCAL GOLD® Fabs for1-2 h at RT. Biotinylated unrelated antigen Transferrin was then addedto the Maxisorp plates to check for biotin binding Fabs (in this casethe HuCAL®-Fab fragments were previously captured via anti-Fd antibody).

Following secondary antibodies were applied for detection: Alkalinephosphatase (AP)-conjugated Streptavidin-AP AffiniPure F(ab′)2 fragment,goat anti-human, was added to the Maxisorp expression plates; anti-HIS6Peroxidase conjugated mouse antibody, Roche, was added to theNeutrAvidin plates and Streptavidin-Alkaline Phosphatase, ZYMED, wasadded to the Maxisorp plates with the biotinylated Transferrin.

For detection of AP-conjugates, fluorogenic substrate AttoPhos (RocheDiagnostics, Mannheim, Germany) and for detection of POD-conjugates,fluorogenic substrate QuantaBlu (Pierce) were used according tomanufacturer's instructions. Fluorescence was measured in a Tecan GENiosPro plate reader.

Using this method it was possible to screen for anti-human C5 Fabs whichrecognize human C5 in solution and to exclude antibodies binding to thebiotin moiety of the target antigens.

3. Determination of Cross-reactivity to Cynomolgus C5

A polyclonal C5-binding antibody (US Biological Cat#C7850-24) was usedto capture cynomolgus C5 from cynomolgus serum.

384 well Maxisorp plates were coated with 20 μl/well of 5 μg/mlpolyclonal C5-binding in PBS and incubated o/n at 4° C. On the next daythe plates were washed 3× with PBST and blocked with 100 μl/well ofdiluent (4% BSA/0.1% Tween20/0.1% Triton-X 100/PBS) for 2 hours at RT.Cynomolgus serum was diluted 1:20 in diluent (4% BSA/0.1% Tween20/0.1%Triton-X 100/PBS) (˜approx. concentration of cynomolgus C5 4 μg/ml) and20 μl/well was added to the 2×PBST washed Maxisorp plates. After 1 hincubation at RT the plates were washed 3×PBST and BEL lysatescontaining Fab fragments or purified Fabs were added and incubated for 1h at RT. The plates were washed again and detection antibodyanti-HIS6-POD (Roche #1965085), was added. POD substrate, BM Blue,soluble, (Roche Applied Science) was added and the reaction was stoppedwith 1M H2SO4. Absorbance was read at 450 nm using the BMG Readerdevice.

Example 4 Affinity Maturation 1. Construction of Affinity MaturationLibraries of Selected C5-Binding Fabs

To increase affinity and biological activity of selected antibodyfragments, L-CDR3 and H-CDR2 regions were optimized in parallel bycassette mutagenesis using trinucleotide directed mutagenesis (see e.g.,Virnekas et al., Nucleic Acids Res. 22:5600-5607 (1994)), while theframework regions were kept constant. Prior to cloning for affinitymaturation, all parental Fab fragments were transferred from thecorresponding expression vector (pMORPH®x9_MH) into the CysDisplay™vector pMORPH®25 via XbaI/EcoRI. pMORPH®25 was created from the HuCALGOLD® display vector pMORPH®23 by removal of one BssHII site interferingwith library cloning for H-CDR2 optimization. For optimizing L-CDR3 ofparental Fabs, the L-CDR3, framework 4 and the constant region of thelight chains (405 bp) of the binders were removed by BpiI/SphI andreplaced by a repertoire of diversified L-CDR3s together with framework4 and the constant domain.

10 parental C5-binding Fabs were divided in 7 pools according todifferent selection criteria and only Fabs with same framework were puttogether: (1) MOR07086; (2) MOR06525+6756 (same framework); (3)MOR06757; (4) MOR06763; (5) MOR07087; (6) MOR07091+7092 (sameframework); (7) MOR07093+7094 (same framework).

Approximately 1.5 μg of the single Fab vector fragment and of the Fabpool were ligated with a 3 to 5-fold molar excess of the insert fragmentcarrying the diversified L-CDR3s. In a second library set, the H-CDR2(XhoI/BssHII) was diversified while the connecting framework regionswere kept constant. In order to monitor the cloning efficiency, theparental H-CDR2 was replaced by a dummy before the diversified H-CDR2cassette was cloned in.

Ligation mixtures of the different libraries were electroporated into E.coli TOP10 F′ cells (Invitrogen) yielding from 2×10⁷ to 2×10⁸independent colonies. The libraries were amplified. For quality control,several single clones per library were randomly picked and sequencedusing primers CFR84 (VL) and OCAL_Seq_Hp (VH).

As described above, seven maturation sub pools were generated and keptseparate during the subsequent selection process.

14 different affinity maturation libraries (one LCDR3 and one HCDR3library for each lead or pool) were generated by standard cloningprocedures and transformation of the diversified clones intoelectro-competent E. coli TOP10F′ cells (Invitrogen). Library sizes weregood, being in the range of 2×10⁷-5×10⁸. Sequencing of randomly pickedclones showed a diversity of 100%. No parental binders but derivativesof all respective parental input binders were found. Finally phages ofall 14 libraries were prepared separately.

TABLE 2 Overview of maturation libraries MOR0 Maturation VH/VL TypeLibrary Size 6757 HCDR2 VH3 3.70 × 10E7 6763 HCDR2 VH3 4.95 × 10E7 7086HCDR2 VH1A 1.58 × 10E8 7087 HCDR2 VH1A 7.85 × 10E7 6525 + 6756 HCDR2 VH55.22 × 10E7 7091 + 7092 HCDR2 VH5 3.51 × 10E7 7093 + 7094 HCDR2 VH2 2.01× 10E7 6757 LCDR3 Vkappa1 1.89 × 10E7 6763 LCDR3 Vlambda2 7.35 × 10E77086 LCDR3 Vlambda3 7.54 × 10E7 7087 LCDR3 Vkappa1 5.46 × 10E7 6525 +6756 LCDR3 Vlambda2 8.50 × 10E7 7091 + 7092 LCDR3 Vlambda3 4.93 × 10E87093 + 7094 LCDR3 Vlambda2 1.33 × 10E8

2. Preparation of Antibody-Phages for Affinity Maturation

The HuCAL® maturation libraries were amplified in 2×YT medium containing34 μg/ml chloramphenicol and 1% glucose (2×YT-CG). After infection withVCSM13 helper phage at an OD600 nm of 0.5 (30 min at 37° C. withoutshaking; 30 min at 37° C. shaking at 250 rpm), cells were spun down(4120×g; 5 min; 4° C.), resuspended in 2×YT/34 μg/ml chloramphenicol/50μg/ml kanamycin/0.25 mM IPTG and grown o/n at 22° C. Phages werePEG-precipitated twice from the supernatant, resuspended in PBS and usedfor the maturation pannings described below.

3. Standard Solution Maturation Panning on Biotinylated C5 Protein

About 10¹² phages rescued from the generated affinity maturationlibraries, as described above, were subjected to pannings performedunder very stringent conditions to select for affinity improved C5specific Fabs.

Solution pannings using the respective phage pools were either performedusing biotinylated human C5 or alternating biotinylated human andcynomolgus C5 proteins. In order to increase panning stringency and toselect for improved off-rates, antigen concentration was decreased andprolonged washing periods were applied (washing conditions are listed inTable 3).

TABLE 3 Increased washing conditions within the selection rounds ofsolution maturation pannings Selection Rd. Washing conditions (modified:stringent) 1st round 4× PBS/0.05% Tween 5 min on rotator 3× PBS/0.05%Tween 15 min on rotator-> transfer magnetic beads with the capturedantigen and phages to a fresh blocked tube 4× PBS quick 3× PBS 5 min onrotator-> transfer magnetic beads with the captured antigen and phagesto a fresh blocked tube 2nd round 3× PBS/0.05% Tween quick 7× PBS/0.05%Tween 15 min on rotator-> transfer magnetic beads with the capturedantigen and phages to a fresh blocked tube 3× PBS quick 7× PBS 15 min onrotator-> transfer magnetic beads with the captured antigen and phagesto a fresh blocked tube 3rd round 5× PBS/0.05% Tween quick 8× PBS/0.05%Tween 15 min on rotator 1× PBS/0.05% Tween o/n on rotator 3× PBS/0.05%Tween quick 6× PBS/0.05% Tween 15 min on rotator -> transfer magneticbeads with the captured antigen and phages to a fresh blocked tube 5×PBS quick 8× PBS 15 min on rotator -> transfer magnetic beads with thecaptured antigen and phages to a fresh blocked tube

Pre-blocked phage (1:2 mixture with 2× Chemiblocker incubated for 1 h atRT) were incubated with low concentration of biotinylated C5 protein for1-2 h at RT. The panning strategy is similar to a standard solutionpanning described above. The phage antigen complex was captured via thebiotin moiety of C5 to pre-blocked Streptavidin magnetic beads 30 min atRT. Beads were then washed more stringently compared to a normalpanning. Elution and amplification of phage was performed as describedabove.

The second and third rounds of selection were performed in an identicalway to the first round, but at higher stringency washing conditions andlower antigen concentrations. For each antibody lead or pool severaldifferent pannings were performed. For each panning strategy differentstringency conditions were applied. Panning strategies are summarized inTable 4.

TABLE 4 Overview of solution maturation pannings 1783 and 1784 onbiotinylated human C5 and biotinylated cynomolgus C5 Panning AntigenAntigen Antigen Panning # Library mode 1st round 2nd round 3rd roundAntigen Conc. Washing 1783.1 MOR06525 + 6756 HCDR2 solution human C5human C5 human C5 50 nM human/ modified 1783.2 MOR07086 HCDR2Streptavidin 5 nM human/ (more 1783.3 MOR06763 HCDR2 beads 0.25 nM humanstringent) 1783.4 MOR07087 HCDR2 1783.5 MOR06525 + 6756 LCDR3 1783.6MOR07086 LCDR3 1783.7 MOR06763 LCDR3 1783.8 MOR07087 LCDR3 1783.9MOR06525 + 6756 HCDR2 solution human C5 cyno C5 human C5 25 nM human/modified 1783.10 MOR07086 HCDR2 Streptavidin 5 nm cyno/ (more 1783.11MOR06763 HCDR2 beads 0.25 nM human stringent) 1783.12 MOR06525 + 6756LCDR3 1783.13 MOR07086 LCDR3 1783.14 MOR06763 LCDR3 1784.1 MOR06757HCDR2 solution human C5 human C5 human C5 50 nM human/ modified 1784.2MOR07091 + 7092 HCDR2 Streptavidin 5 nM human/ (more 1784.3 MOR07093 +7094 HCDR2 beads 0.25 nM human stringent) 1784.4 MOR06757 LCDR3 1784.5MOR07091 + 7092 LCDR3 1784.6 MOR07093 + 7094 LCDR3 1784.7 MOR06757 HCDR2solution human C5 cyno C5 human C5 25 nM human/ modified 1784.8MOR07091 + 7092 HCDR2 Streptavidin 5 nM cyno/ (more 1784.9 MOR07093 +7094 HCDR2 beads 0.25 nM human stringent) 1784.10 MOR07087 HCDR2 1784.11MOR06757 LCDR3 1784.12 MOR07091 + 7092 LCDR3 1784.13 MOR07093 + 7094LCDR3 1784.14 MOR07087 LCDR3

After maturation pannings, the enriched phagemid pools were sub-clonedinto pMORPH®x9_MH expression vector.

4. Cross-Combination of Optimized VL (L-CDR3) with Optimized VH (H-CDR2)

For further improvement of affinity and potency, the independentlyoptimized heavy and light chains from matured antibodies, derived fromthe same parental clone, were combined (see e.g., Rauchenberger et al.,J. Biol. Chem. 278:38194-38205 (2003); Chen et al., J. Mol. Biol.293:865-881 (1999); and Schier et al., J. Mol. Biol. 263:551-567(1996)). This procedure, called cross-cloning, was applied for bindersderiving from the same parental clones.

5. Affinity Screening and Maturation Panning Outcome

A total of 2640 clones derived from all pannings were screened asbacterial lysates for improved affinities on human C5. Preliminaryaffinities were estimated by solution equilibrium titration (SET). Basedon their estimated affinities, clones derived from each parental Fab orFab pools were sequenced. Table 5 shows number of sequenced clones andnumber of obtained unique sequences for each panning condition.

TABLE 5 Overview of affinity improved clones selected for sequenceanalysis Sequenced Unique Parental/Maturation Antigen clones SequencesParental of unique MOR06525 + 6756 HCDR2 hu/hu/hu 10 9 6525 MOR07086HCDR2 hu/hu/hu 10 4 7086 MOR06763 HCDR2 hu/hu/hu 22 10 6763 (8× ), 7086(2× ) MOR07087 HCDR2 hu/hu/hu 10 4 7087 MOR06757 HCDR2 hu/hu/hu 10 0MOR07091 + 7092 HCDR2 hu/hu/hu 24 7 7092 MOR07093 + 7094 HCDR2 hu/hu/hu10 10 7093 MOR06525 + 6756 LCDR3 hu/hu/hu 20 5 6756 MOR07086 LCDR3hu/hu/hu 10 5 7086 MOR06763 LCDR3 hu/hu/hu 10 8 7086 MOR07087 LCDR3hu/hu/hu 6 1 7086 MOR06757 LCDR3 hu/hu/hu 16 0 MOR07091 + 7092 LCDR3hu/hu/hu 6 6 7091 (1× ), 7092 (5× ) MOR07093 + 7094 LCDR3 hu/hu/hu 10 97094 MOR06525 + 6756 HCDR2 hu/cyno/hu 10 8 6525 MOR07086 HCDR2hu/cyno/hu 10 6 7086 MOR06763 HCDR2 hu/cyno/hu 22 5 6763 MOR06757 HCDR2hu/cyno/hu 15 2 6757 MOR07091 + 7092 HCDR2 hu/cyno/hu 15 6 7091 (3× ),7092 (3× ) MOR07093 + 7094 HCDR2 hu/cyno/hu 10 10 7093 MOR07087 HCDR2hu/cyno/hu 10 6 7087 (5× ), 7086 (1× ) MOR06525 + 6756 LCDR3 hu/cyno/hu12 0 MOR07086 LCDR3 hu/cyno/hu 10 1 7086 MOR06763 LCDR3 hu/cyno/hu 10 0MOR06757 LCDR3 hu/cyno/hu 9 1 7094 MOR07091 + 7092 LCDR3 hu/cyno/hu 11 97091 (6× ), 7092 (3× ) MOR07093 + 7094 LCDR3 hu/cyno/hu 10 7 7094MOR07087 LCDR3 hu/cyno/hu 10 0 Sum 338 139

6. Sequence Analysis and Selection of Affinity Optimized Fabs forProtein Production

A very good diversity was maintained by recovering derivatives of all 10parental Fabs. The nucleotide sequences of the heavy chain (VH) for 188HCDR2 improved clones and the light chain (VL) variable regions for 150improved LCDR3 clones were determined. 87 unique HCDR2 and 52 uniqueLCDR3 sequences were selected for a detailed analysis of sequencediversity within the matured CDRs. Fabs containing possibleglycosylations sites in the CDRs were omitted from furthercharacterizations.

The VH and VL sequence analysis and affinity data showed that all 10parental Fabs yielded affinity-improved successors. Parental FabsMOR06525, MOR06757, MOR06763, MOR07087 and MOR07094 yielded only HCDR2improved clones and parentals MOR06756 and MOR07093 yielded only LCDR3improved clones. MOR07086, MOR07091 and MOR07092 had matured clones forboth VH and VL. This later allowed cross-cloning of VH and VL maturedchains. From all data, 60 clones with best affinity and highestdiversity in the matured CDRs were selected for Fab expression. SelectedVH and VL amino acid, as well as nucleotide sequences, are listed inTable 1.

Example 5 IgG Conversion

1. Conversion into Human IgG2 Format

In order to express full length immunoglobulin (Ig), variable domainfragments of heavy (VH) and light chains (VL) were subcloned from thepMORPH®x9_MH Fab expression vectors into pMORPH®2_h_Ig vector series forhuman IgG2. Restriction enzymes MfeI, and BlpI were used for subcloningof the VH domain fragment into pMORPH®2_h_IgG2. Subcloning of the VLdomain fragment into pMORPH®2_h_Igκ was performed via the EcoRV andBsiWI sites, whereas subcloning into pMORPH®2_h_Igλ2 was done usingEcoRV and HpaI.

All ten parental Fabs (MOR06525, 6756, 6757, 6763, 7086, 7087, MOR07091,7092, 7093 and 7094) were converted into human IgG2. The IgGs were alsoexpressed.

2. Conversion into Human IgG1AA Format

In order to express full length immunoglobulin, variable domainfragments of Fab heavy (VH) and light chains (VL) were subcloned fromthe Fab expression vectors into IgG1 expression vectors. Restrictionenzymes MfeI, and BlpI were used for subcloning of the VH domainfragment into pMORPH®2_h_IgG1AA, in which leucines at positions 234 and235 were mutated to alanines to abrogate FcRγ binding and attenuateeffector functions. The restrictions enzymes EcoRV and HpaI were used tosubclone of the VL domain fragment into pMORPH®2_h_Igλ2.

Following matured Fabs with desired profile were subcloned into humanIgG1AA format: MOR07832, 7834, 7872, 7876, 7829, 7871, 7865, 7873, 7830,7878, 7910. Cross-cloning on IgG level was achieved by transfectingcells with combinations of light and heavy chain constructs. Forexample, MOR08114 was the product of the germlined heavy chain fromMOR07829 and the germlined light chain from MOR07871. Table 6 summarizesthe most relevant cross-cloned germlined IgGs.

TABLE 6 Overview of most relevant cross-cloned germlined IgGs maturedCDRs MOR0 VH/VL VH/VL matured Nr. germlined VH VL matured VH VL format8114 yes 7829 7871 7091/HCDR2 7091/ huIg1AA LCDR3 8125 yes 7091 7873 —7091/ huIg1AA LCDR3 8126 yes 7829 7873 7091/HCDR2 7091/ huIg1AA LCDR38127 yes 7830 7873 7091/HCDR2 7091/ huIg1AA LCDR3 8128 yes 7092 7878 —7092/ huIg1AA LCDR3 8129 yes 7909 7092 7092/HCDR2 — huIg1AA 8130 yes7909 7878 7092/HCDR2 7092/ huIg1AA LCDR3 8131 yes 7910 7092 7092/HCDR2 —huIg1AA 8132 yes 7910 7878 7092/HCDR2 7092/ huIg1AA LCDR3

3. Transient Expression and Purification of Human IgG

Eukaryotic HKB11 and HEK293 cells were transfected with an equimolarratio of IgG heavy and light chain expression vector DNA. Cell culturesupernatant was harvested at 3 or 7 days post transfection and subjectedto standard protein A affinity chromatography (rProteinA FF or MabSelectSURE, GE Healthcare). As not otherwise stated, buffer exchange wasperformed to 1× Dulbcecco's PBS (pH 7.2, Invitrogen) and samples weresterile filtered (0.2 μm). Purity of IgG was analyzed under denaturing,reducing and non-reducing conditions in SDS-PAGE or by using AgilentBioAnalyzer and in native state by HP-SEC.

Example 6 Germlining

IgG constructs were germlined via site-directed mutagenesis usingQuickChange® Site-Directed Mutagenesis Kit (Stratagene). The N-terminalDI of MOR08111 Vλ2 were changed to ES to match human germline sequenceas well as to avoid a terminal Q (N-terminal Q can form pyroglutamine).N-terminal DI of MOR08110 Vλ3, MORO8113 Vλ3, and MOR08114 Vλ3 weregermlined to SY, the most commonly found sequence in human λ3 genes.N-terminal QVQ of MOR08111 VH2 was germlined to EVT to match a λ2 geneand avoid terminal Q. N-terminal Q in MOR08109 VH5, MOR08110 VH5,MOR08113 VH5 and MOR08114 VH5 was also mutated to E.

Framework sequences for MOR08109 Vλ3 were synthesized to match the humanλ3j gene and cloned into the expression vector using NheI and HpaIrestriction sites. Sequence alignments of the antibodies variabledomains with their respective closest related human germline sequencesare shown in FIG. 1.

Example 7 Affinity Determination 1. Kon/Koff and K_(D) Determination ofAnti-Human C5 Antibodies Using Surface Plasmon Resonance (Biacore)

It was determined that anti-Fab antibodies used to immobilize Fabs tothe Biacore chip were influencing differently the binding affinity ofeach Fab for human C5, thus making the comparison of the Fabs to eachother difficult. Biacore analysis was performed on IgG antibodies.

A CM4 chip was coated with 50 μg/ml goat anti-human Fc antibody(500-2000 RU) in 10 mM acetate buffer, pH 4.5, using standard EDC-NHSamine coupling chemistry. Each anti-human C5 IgG was captured on thechip in HBS-EP buffer at constant flow rate of 10 μl/min for a contacttime leading to a ligand density around 20 RU. After capturing theanti-hu C5 IgG, different concentrations of human or cynomolgus C5, inthe range between 0.156 nM to 2.5 nM, were injected. Each cycle wascompleted with two regeneration steps with phosphoric acid. All runningconditions were carried out at 25° C. in 1×HBS-EP buffer. The resultingsignals were adjusted by double referencing, subtracting the refractionindex values from the reference flow cell and the binding step with noanalyte. Data were collected at 10 Hz and analyzed using the BiacoreT100 Evaluation Software Version 1.1 (GE). This program uses a globalfitting analysis method for the determination of rate and affinityconstants for each interaction.

The specificity of the antibodies were measured. Preferably, the Kon andKoff values for binding to human and cynomolgus C5 are as follows:Kon>1×10⁵, Koff<1×10⁴). These measurements were performed in Biacore forthe germlined IgGs and resulting data are listed in Table 7.

TABLE 7 K_(D), Kon and Koff values of the germlined IgGs determined inBiacore antiC5 final IgG C5 sample ka [1/Ms] kd [1/s] KD [pM] MOR08109huC5 2.13E+06 2.56E−05 12 cynoC5 1.23E+06 4.49E−05 37 MOR08110 huC54.15E+06 4.69E−05 12 cynoC5 1.81E+06 9.24E−05 60 MOR08111 huC5 1.00E+063.07E−05 31 cynoC5 8.91E+05 1.28E−04 144 MOR08113 huC5 2.51E+06 6.77E−0528 cynoC5 1.53E+06 1.27E−04 83 MOR08114 huC5 2.09E+06 3.12E−05 15 cynoC51.06E+06 3.13E−05 31 5G1.1 huC5 1.29E+06 7.22E−05 56

2. Determination of Picomolar Affinities Using Solution EquilibriumTitration (SET) for Purified Fabs or Fabs Bacterial Lysates (Meso ScaleDiscovery (MSD))

For K_(D) determination by solution equilibrium titration (SET), monomerfractions (at least 90% monomer content, analyzed by analytical SEC;Superdex75, Amersham Pharmacia) of Fab protein were used. Affinitydetermination in solution was basically performed as described in theliterature (Friguet et al., J. Immunol. Methods 77:305-319 (1985)). Inorder to improve the sensitivity and accuracy of the SET method, it wastransferred from classical ELISA to ECL based technology (Haenel et al.,Anal Biochem 339:182-184 (2005).

1 mg/ml goat-anti-human (Fab)₂ fragment specific antibodies (Dianova)were labelled with ECL Sulfo-TAGTM NHS-Ester (Meso Scale Discovery,Gaithersburg, Md., USA) according to manufacturer's instructions.Experiments were carried out in polypropylene microtiter plates and PBSpH 7.4 with 0.5% BSA and 0.02% Tween 20 as assay buffer. Unlabelledantigen was diluted in 2^(n) series, starting with a concentration atleast 10 times higher than the K_(D). Wells without antigen were used todetermine Bmax values; wells with neither antigen nor Fab were used todetermine background. After addition of e.g. 10 pM Fab (finalconcentration in 60 μl final volume), the mixture was incubated overnight at RT. The applied Fab concentration was similar to or below theexpected K_(D).

Streptavidin MSD plates were coated with 0.2 μg/ml biotinylated human C5(30 μl/well) and blocked with 5% BSA in PBS. Subsequently theequilibrated samples were transferred to those plates (30 μl per well)and incubated for 20 min. After washing, 30 μl/well of the ECL Sulfo-taglabeled detection antibody (goat anti-human (Fab)₂) in a final dilutionof 1:1500 was added to the MSD plate and incubated for 30 min on anEppendorf shaker (700 rpm).

After washing and adding 30 μl/well MSD Read Buffer T with surfactantElectrochemiluminescence signals were detected using a Sector Imager6000 (Meso Scale Discovery, Gaithersburg, Md., USA).

Data were evaluated with XLfit (IDBS) software applying customizedfitting models. For data evaluation i.e. K_(D) determination of Fabmolecules the following fit model was used (model of Abraham et al 16,modified according to et al., 200515):y=Bmax−(Bmax/(2*cFab)*(x+cFab+KD−sqrt((x+cFab+KD)*(x+cFab+KD)−4*x*cFab)));cFab: applied Fab concentration; x: applied total soluble antigenconcentration (binding sites); sqrt: square root. Using the assayconditions described above (monomeric) affinities for theaffinity-optimized C5-binding Fabs were determined in solution.

Parental Fabs

In order to further characterize the C5-binding antibodies, affinity ofthe parental Fabs to human C5 was determined. Because characterizationfocus was on efficacy in hemolytic assays, affinity measurements weredone only for the most relevant Fabs. For a reliable determination ofmonovalent affinities only Fab batches were used for measurements whichshowed >90% monomeric fraction in a qualitative size exclusionchromatography.

Affinities of the 10 parental Fabs which entered affinity maturation aresummarized in Table 8. Affinities ranged from 72 pM to 3.7 nM.

TABLE 8 Affinities of the 10 parental Fabs determined in SET SET MOR0Number KD [pM] 6525 72 6756 1521 6757 1186 6763 820 7086 108 7087 37937091 324 7092 229 7093 576 7094 1364 3207 no binding (negative control)(n = 1)

Matured Fabs

Monovalent affinities of the purified Fabs to human C5 were measured inSET. Affinities were in the low pM range and best affinities wereobtained for derivatives of MOR07086, 7091, 7092 and 7093. Subsequentlyaffinity measurements of these derivatives to cynomolgus C5 showedaffinities in the mid to low pM range.

The affinity maturation process was very successful resulting in arepertoire of binders with markedly improved affinity. Table 9summarizes affinities to human and cynomolgus C5 of the best improvedbinders. Certain Fabs have K_(D) to human C5≦30 pM and to cynomolgusC5≦150 pM.

TABLE 9 Overview of affinities to human and cynomolgus C5 for the bestaffinity improved Fabs

3. K_(D) Determination of IgG Molecules Using Solution EquilibriumTitration (SET)

Affinities of the germlined IgGs (human IgG1AA format) to human andcynomolgus C5 were determined in SET as described below. Similar datasets between two independent measurements showed higher affinities ofthe lead IgGs to human C5 than reference IgG 5G1.1 (see U.S. Pat. No.6,355,245). Final IgGs had affinities for human C5 ranging from 1 to 14pM and affinities to cynomolgus C5 ranging from 3 to 29 pM.

TABLE 10 K_(D) values determination for the final lead IgGs (humanIgG1AA format) in SET 1^(st) measurement 2^(nd) measurement human C5cyno C5 human C5 cyno C5 KD [pM] KD [pM] KD [pM] KD [pM] hu MOR08109 413 2 6 IgG1AA MOR08110 7 18 3 8 germlined MOR08111 5 14 3 17 MOR08113 1429 8 16 MOR08114 1 5 2 4 hu IgG2/4 5G1.1 24 no binding 19 no binding(reference IgG)

For K_(D) determination by solution equilibrium titration (SET), monomerfractions of IgG protein were used (at least 90% monomer content,analyzed by analytical SEC MALS; Tosoh TSKgel G3000SWXL, Wyatt TreosminiDAWN). Affinity determination in solution was basically performed asdescribed in the literature (Friguet et al., J. Immunol. Methods77:305-319 (1985)). In order to improve the sensitivity and accuracy ofthe SET method, it was transferred from classical ELISA to ECL basedtechnology (Haenel et al., Anal Biochem 339:182-184 (2005)).

1 mg/ml goat-anti-human (Fab)₂ fragment specific antibodies (Dianova)were labelled with ECL Sulfo-TAGTM NHS-Ester (Meso Scale Discovery,Gaithersburg, Md., USA) according to the manufacturer's instructions.The experiments were carried out in polypropylene microtiter plates andPBS pH 7.4 with 0.5% BSA and 0.02% Tween 20 as assay buffer. Unlabeledantigen was diluted in 2n or 1.75n series, respectively, starting with aconcentration at least 10 timer higher than the K_(D). Wells withoutantigen were used to determine Bmax values; wells containing neitherantigen nor IgG were used to determine background. After addition ofe.g. 10 pM IgG (final concentration in 60 μl final volume), the mixturewas incubated over night at RT. The applied IgG concentration wassimilar to or below the expected K_(D).

Streptavidin MSD plates were coated with 0.2 μg/ml biotinylated human C5(3 μl/well) and blocked with 5% BSA in PBS. Subsequently theequilibrated samples were transferred to those plates (30 μl per well)and incubated for 20 min. After washing, 30 μl/well of the ECL Sulfo-taglabeled detection antibody (goat anti-human (Fab)₂) in a final dilutionof 1:1500 was added to the MSD plate and incubated for 30 min on anEppendorf shaker (700 rpm).

Electrochemiluminescence signals were detected after washing and adding30 μl/well MSD Read Buffer T with surfactant using a Sector Imager 6000(Meso Scale Discovery, Gaithersburg, Md., USA).

Data were evaluated with XLfit (IDBS) software applying customizedfitting models. For data evaluation i.e. K_(D) determination of IgGmolecules the following fit model for IgG was used (modified accordingto Piehler et al., 199717):y=Bmax/(cIgG/2)*(cIgG/2−((x+cIgG+KD)/2−((x+cIgG+KD)̂2/4−x*cIgĜ0.5)̂2/(2*IgG));cIgG=applied IgG concentration, complete molecule (not binding sites);x=applied total soluble antigen concentration (binding sites); sqrt:square root.

Example 8 Characterization by Hemolytic Assays

The hemolytic assay is a basic functional assay that tests forcomplement activation and has been used to evaluate the ability ofanti-human C5 mAbs and Fab molecules to block lysis of red blood cells(RBCs) by complement pathways (see e.g., Evans et al., Mol. Immunol. 32:1183-1195 (1995); Thomas et al., Mol Immunol 33:1389-1401 (1996); Rinderet al., J Clin Invest 96:1564-1572 (1995)). Briefly, for classicalpathway assays, sensitized red blood cells are used as targets for lysisby complement proteins present in serum. This assay is of interest forthe characterization and screening of high-affinity anti-human C5 mAbs.

1. Classical Pathway

The desired number of chicken red blood cells was washed four times withcold gelatin veronal buffer (GVB++) and resuspended to 5×10⁷ cells/ml.To sensitize the cells rabbit anti-chRBC IgG was added to RBC cellsuspension to a final concentration of 1 μg/ml IgG. After 15 minutesincubation on ice, the sensitized chRBCs were centrifuged, washed twicewith GVB++ and diluted to 8.33×10⁷ cells/ml.

Round-bottom 96 well plates were used for hemolytic assay. Antibodieswere diluted in GVB++ buffer and added to the wells (when calculatingthe required concentration of C5-binding Abs, it was considered that thesample will be diluted two-fold when serum is added). 50 μl of 40% humanserum (diluted in GVB++) was added to 50 μl antibody dilutions,resulting in a final serum assay concentration of 20%.

The control and blank wells were prepared as described here: controlwells: i) 0% lysis control→100 μl GVB++, ii) 100% lysis control→100 μl0.1% NP-40, iii) 20% serum control→100 μl of 20% serum (0% Ab control).blank wells: i) 20% serum blank→100 μl 20% serum, ii) GVB++ blank→100 μlGVB++, iii) NP-40 blank→100 μl 0.1% NP-40.

2.5×10⁶ (30 μl) sensitized chRBCs/well were added to all sample andcontrol wells. To the blank wells PBS was added instead of cells. Assayplate was incubated 30 min at 37° C., centrifuged (2.000 rpm, 5 min) and85 μl supernatant was transferred to a new, flat-bottomed 96-well plate.The new plate was centrifuged (2.000 rpm, 3 min) to get rid of anybubbles. Hemoglobin release was measured by reading absorbance at 415nm. Percentage of hemolysis was calculated with respect to the controland blank wells using the following calculation algorithms:

${\% \mspace{14mu} {Hemolysis}} = {100 \times \frac{{ODsample} - {ODnegativecontrol}}{{ODpositivecontrol} - {ODnegativecontrol}}}$whereODsample = ⌊AverageOD_(sample)⌋ − [AverageOD_(20%  Serum  Blank)]ODnegativecontrol = ⌊AverageOD_(0%  Lysis)⌋ − [AverageOD_(GVB + +Blank)]ODpositivecontrol = ⌊AverageOD_(100%  Lysis)⌋ − [AverageOD_(NP − 40  Blank)]

Using this procedure, anti human-C5 antibodies which were able toinhibit red blood cell lysis could be identified. To screen forcross-reactivity to cynomolgus C5, the classical pathway was performedusing 5% cynomolgus serum.

2. Alternative Pathway

Hemolytic assays undergoing the alternative pathway were done in asimilar way to the classical pathway hemolytic assays. In thealternative pathway RBCs cells from rabbit were used and there was noneed to sensitize the cells. The rabbit RBCs are different from chickenRBCs in that they are sensitive to lysis caused by the complementalternative pathway.

The working buffer was GVB++ supplemented with 10 mM EGTA and 5 mM Mg++,since the C5 convertase of the alternative pathway is Mg++dependent andthe C5 convertase of the classical pathway is Ca++ dependent.

Hemolytic assays of the alternative pathway were run with: i) 20% humanserum, ii) 100 pM human C5 added to 20% human C5-depleted serum, iii)0.025% cynomolgus serum added to 20% human C5-depleted serum, iv) 100 pMcynomolgus C5 added to 20% human C5-depleted serum, v) 10% cynomolgusserum. These settings were used to screen for antibodies with highaffinity to the human and cynomolgus C5 proteins which were able toinhibit very effectively the red blood cell lysis induced by thealternative complement pathway.

3. Hemolytic Assays with Parental Fabs

Hemolytic assays were used as a basic bio-functional assay to evaluatethe ability of anti-human C5 mAbs to block complement mediated lysis ofred blood cells. C5 convertase cleaves C5 into C5a peptide and C5bfragment, that is subsequently incorporated into the membrane-attackcomplex (MAC), which leads to cell lysis. C5 convertase of the classicalpathway, formed by a C3bC4bC2a complex has a different structure thanthe C5 convertase of the alternative pathway which is formed by aC3bC3bBb complex. HuCAL GOLD® generated antibodies should be inhibitoryin both classical and alternative pathway, but with focus on thealternative pathway because mainly the alternative pathway (factor H,factor B and factor H-related genes) is implicated in AMD.

The classical and alternative pathway assays were performed with 20%human serum (˜80 nM C5). To increase sensitivity of alternative pathwayassays, new assay formats were developed. 10-100 pM purified human C5 or0.025% cynomolgus serum (˜100 pM cynomolgus C5) were added to humanC5-depleted serum (but containing all other serum and complementcomponents).

FIG. 2 shows that considerable hemolysis could be observed between 10and 100 pM purified human C5 added to human C5-depleted serum.Cynomolgus serum was added to human C5-depleted serum to test forcross-reactivity. FIG. 3 shows that 0.025% of cynomolgus serum (˜100 pMC5) added to human C5 depleted serum restores hemolytic activity.

Classical Pathway

First Fab selection was done in the classical pathway (20% human serum).Approximately half of the 61 purified parental Fabs were weak to stronginhibitors of the classical pathway. IC50 values of the best inhibitoryFabs were between 35 and 900 nM.

Assays were done showing congruent results (as shown in FIG. 4). %hemolysis was calculated with respect to the control and blank wells.Fab inhibition of cell lysis was compared to a maximum lysis caused by20% human serum (=100%). An irrelevant human Fab (hen egg white lysozymebinder MOR03207) was used as negative control and anti-human C5 IgGmonoclonal antibody (Quidel) as positive control. FIG. 4 show an examplewith the best inhibitory Fabs.

Alternative Pathway

Fabs which showed inhibitory activity in the classical pathway werefurther evaluated in the alternative pathway. Hemolytic assays were runwith 100 pM purified human C5 or 0.025% cynomolgus serum added to humanC5-depleted serum. IC₅₀ values for the human alternative assays werebetween 0.1 and 90 nM (examples of assays with the most relevant Fabsare shown in FIG. 5.

The positive control of the classical pathway (anti-human C5 antibody,Quidel) was not inhibitory in the alternative pathway. Therefore ananti-complement factor P antibody (Quidel) was used as positive control.As shown in FIG. 5, MOR07086 had best inhibitory activity and NVS datarevealed a better potency than for the reference antibody 5G1.1.

To test for cynomolgus cross-reactivity, hemolytic assays of thealternative pathway were performed with 0.025% cynomolgus serum added tohuman C5-depleted serum. A comparison to 5G1.1 was not possible, since5G1.1 does not recognize cynomolgus C5. The anti-Factor P antibody wasused as positive control. Results of assays revealed IC₅₀ values between0.1 and 400 nM for the best inhibitory Fabs. Again, MOR07086 showed bestpotency (shown in FIG. 6).

A consistent inhibitory activity of the Fabs was noticed in bothclassical and alternative pathway. Table 11 below summarizes the resultsof hemolytic assays for the most relevant 22 Fabs. To have a reliablecomparison between different experiments, lysis caused by 20% humanserum was normalized to 100%.

TABLE 11 Summary of hemolytic assays with the most relevant Fabs MORIC50 [nM] NVS IC50 [nM] AP (0.1 nM C5) AP (0.025% cyno AP (0.1 nM C5) AP(0.025% cyno CP [human] [human] serum) [cyno] CP [human] [human] serum)[cyno] MOR-Nr normalized normalized normalized normalized normalizednomialized 6525 190 15 11 185  7 5 6756 320 80 400 225 70 2500 6757 50090 30 305 130  25 6763 250 45 110 195 20 360 6764 n.t. 50 n.t. n.t 2530% inh 6776 >4000  40 n.t.  20* 50% inh 6952 90 20 >1000 110 15 2006961 100 25 600 85 15 30 7081 180 5 40% inh 170  3 10 7082 70 2.5 1 90 1 1 7083 100 30 300 140 10 5 7084 120 10 1.2 160  5 1.5 7086  350.2/0.2 0.2/0.4 85   0.1 0.1 7087 >4000  50 100 775 10 1 7088 110 15 230130  5 15 7089 150 75 900 250 20 50 7090 105 20 10 120 10 1 7091  82 740 110  3 4 7092 100 1 1.5 90   0.5 1.5 7093 >4000  7 190 230  5 15 7094770 40 190  7095*  120* 0.5** 1.3** n.t. *not pure as MH **as pMx9_FS4. Hemolytic Assays with Matured Fabs

Classical Pathway (1) Classical Pathway Using 20% Human Serum

Matured Fabs were tested in the classical pathway with 20% human serum.Derivatives of MOR07086, 7091, 7092 and 7093 showed highest potency(IC50 values in the low nM range). Descendants of MOR07091, 7092 and7093 showed strongly improved potency. FIG. 7 shows examples ofhemolytic assays with derivatives of MOR07086, 7091, 7092 and 7093.

(2) Classical Pathway Using 5% Cynomolgus Serum

Assays of the complement pathway were also run in the presence of 5%cynomolgus serum in order to test for cross-reactivity. Derivatives ofMOR07086, 7091, 7092 and 7093 could very effectively inhibit red bloodcell lysis. The negative control, MOR03207 (anti-lysozyme Fab), had noimpact on the complement pathway. Results of these assays are shown inFIG. 8.

Alternative Pathway (1) Alternative Pathway Using 100 pM Human C5

Matured Fabs were tested in the alternative pathway hemolytic assay with100 pM human C5. Some derivatives of MOR06525, 6757, 6763, and 7087showed potency improvement compared to their parentals. MOR07086-,7091-, 7092-, 7093-, and 7094-derived Fabs showed highest potency (IC50values in the low nM range). Descendants of MOR07091, 7092, 7093, and7094 showed highly improved potency, many of which are more potent thanreference antibody 5G1.1. FIG. 9 shows examples of hemolytic assayresults for the affinity matured Fabs and 5G1.1.

(2) Alternative Pathway Using 20% Human Serum

Matured Fabs were tested in the alternative pathway hemolytic assay with20% human serum. MOR07086-, 7091-, 7092- and 7093-derived Fabs showedbest inhibitory activity. Many of these Fabs had better inhibitoryactivity than 5G1.1. FIG. 10 shows examples of hemolytic assay resultsfor the affinity matured Fabs and reference antibody 5G1.1.

(3) Alternative Pathway Using 100 pM Cynomolgus C5

Matured Fabs were tested in the alternative pathway hemolytic assayusing 100 pM cynomolgus C5 added to 20% human C5-depleted serum.MOR07091-, 7092- and 7093-derived Fabs showed best inhibitory activity;5G1.1 does not crossreact with cynomolgus C5. FIG. 11 shows examples ofhemolytic assay results for the affinity matured Fabs.

5. Hemolytic Assays with Germlined IgGs (Human IgG1AA Format)

Classical Pathway (1) Classical Pathway Using 20% Human Serum

Classical pathway assays using 20% human serum were run at MOR. IC50values of the final germlined hu IgGAA—MOR08109, 8110, 8113, 8114— werebetter or similar to reference IgG 5G1.1 (see FIG. 12).

(2) Classical Pathway Using 5% Cynomolgus Serum

A comparison to 5G1.1 in the classical pathway using 5% cynomolgus serumwas not applicable, since this reference antibody does not recognizecynomolgus C5. The final germlined IgGs could completely inhibit lysisof the red blood cells induced by cynomolgus serum except MOR08111. Dataare shown in FIG. 13.

Alternative Pathway (1) Alternative Pathway Using 100 pM Human C5

The germlined IgGs were tested in the alternative pathway hemolyticassay using 100 pM human C5. All antibodies showed potent inhibitoryactivity with IC50 values between 28 and 128 pM (with the exception ofMOR08111, see FIG. 14), all were equal to or better than 5G1.1. FIG. 14shows examples of hemolytic assay results for the IgGs.

(2) Alternative Pathway Using 20% Human Serum and C5a Generation ELISA

The germlined IgGs were also tested in the alternative pathway hemolyticassay with 20% human serum. The majority of the antibodies testedachieve complete inhibition with IC50 values lower than 80 nM. Referenceantibody 5G1.1 does not fully inhibit hemolysis in this assay. FIG. 15shows examples of hemolytic assay results for the IgGs. Inhibition ofC5a generation by the final IgGs was similar to 5G1.1 (IC50 values inthe low nM range).

(3) Alternative Pathway Using 100 pM Cynomolgus C5

Hemolytic assays of the alternative pathway in 20% human C5-depletedserum were reconstituted with 100 pM cynomolgus C5. Potency of thegermlined final candidates against cynomolgus C5 was within 5-fold ofthat for human C5 (IC50 values in the low pM range).

(4) Alternative Pathway Using 10% Cynomolgus Serum

In hemolytic assays of the alternative pathway using 10% cynomolgusserum ([C5]˜40 nM) the potency of the germlined candidates was similarto the potency in human serum (success criterion was to have a potencynot more than 5-fold weaker than for the functional assay using humanC5).

Example 9 C5a Generation ELISA

C5a-des-Arg ELISA was developed to measure C5a generation duringhemolysis to confirm that antibodies that were inhibitory in thehemolytic assay also inhibited cleavage of C5 into C5a and C5b.

A Maxisorp plate was coated with 100 μl/well mouse anti-humanC5a-des-Arg (US Biologics) at 1 μg/ml in coating buffer (bicarbonate pH9.5-9.8) and was incubated overnight at 4° C. After washing 3× withPBST, the plate was blocked with 300 μl/well diluent (Synblock, AbDSerotec) for 2 hours at room temperature. After aspirating the blockingsolution, 100 μl samples or standards diluted with diluent wereincubated for 1 hour at room temperature.

Standards were prepared as follows: start was at 20 ng/ml standard(rC5a-des-Arg) and 1:4 serial dilutions were prepared for a 7-pointcurve. Samples of hemolytic assays were diluted 1:5 in diluent(hemolytic assay supernatants should be stored at −80° C. until used inC5a ELISA). In between the plate was washed 3× with PBST. 100 μl/well of0.4 μg/ml detection antibody (biotin-goat anti-human c5a, R&D Systems)diluted in diluent was added and after 1 hour incubation at roomtemperature, 100 μl/well Strep-HRP (poly-HRP streptavidin) diluted1:5000 in HRP diluent (poly-HRP diluent) was added for 30 minutes. Afterwashing 4× with PBST, 100 μl/well TMB Substrate (Ultra TMB substratesolution) was added for 5-10 minutes. Reaction was stopped with 50μl/well stop solution (2NH2SO4). Absorbance was read (A450-A570) anddata were analyzed using SoftMax Pro.

Matured Fabs were tested for C5a generation during hemolysis to confirmthat inhibitory activity was due to blocking C5 cleavage into C5a andC5b. The supernatants from hemolytic assays in 20% human serum were usedfor quantifying the C5a formation.

All Fabs tested brought C5a levels down to baseline. FIG. 16 showsexamples of C5a ELISA results.

Example 10 Specificity ELISA on Human C3, C4, C5 and Cynomolgus C5

All purified Fabs were analyzed in a solution ELISA (method describedabove) for binding to human C3, C4 and C5. Fabs were incubated withbiotinylated antigen on a Neutravidin plate and detected via thehistidin tag.

Improved binding was seen for almost all matured Fabs compared to theirrespective parental. No binding to the counter targets human C4 and C3was detected up to 100 nM Fab. These results hit the success criteriafor specificity: binding to human and cynomolgus C5 and no binding tohuman complement proteins C3 and C4. Examples for derivatives ofparental Fab MOR07091 are shown in FIG. 17.

Example 11 Serum Stability Assays

Retained binding activity to human C5 in a binding assay at 50% humanserum of C5-binding antibodies was determined as described below.

Antibodies (Fab format) were incubated up to 8 h at 37° C. with 100%human C5-depleted serum or with PBST/0.5% BSA (positive control). Wellsof a blocked polypropylene plate were used for incubation to ensure nobinding of the antibodies to the surface over the long incubation time.Samples were collected at different time points and stored at −20° C.

Samples were tested in a solution ELISA on NeutrAvidin plates to checkbinding ability to human C5. To the NeutrAvidin plates, which wereblocked o/n with 1× ChemiBlocker-PBST. 20 μl of serial dilutions of thedifferent collected samples were added. First dilution of the sampleswas 1:2 (final serum concentration 50%), followed by 1:3 dilutionssteps. After 1 h incubation the plate was washed 3× with PBST and 20 μlbiotinylated human C5 was applied to a concentration of 2.5 μg/ml. After1 h plate was washed again 5× with PBST (0.05% Tween) and anti-HIS6-PODdetection antibody for Fabs was added.

Fluorescence of the substrate (Quanta Blue or AftoPhos) was measuredafter 5-10 min and retained binding activity was calculated comparedwith the respective maximum signal (antibody incubated with PBST/0.5%BSA).

One of the “must” criteria for the C5-binding antibodies is to retain75-80% of binding activity in human serum i) in a functional assay at10% serum and ii) in a binding assay at 50% serum. Because hemolyticassays were run in the presence of 20% serum it was only necessary toshow retained binding in a binding assay at 50% serum.

Therefore matured final Fabs were incubated with 100% human C5-depletedserum at 37° C. for 8 h. Samples were collected at different time pointsand tested for binding to human C5 in a solution ELISA. Fab+serumsamples used for ELISA were diluted to a concentration of 50% serum+10nM Fab.

FIG. 18 illustrates the results of the final C5-binding final antibodiesin the Fab format. 70-93% of the binding activity was retained after an8 hour incubation time at 37° C. in 50% serum compared to incubation inPBS.

Example 12 Characterization by Epitope Binning

This procedure was used to group anti-human C5 Fabs into differentepitope bins binding to the same or an overlapping epitope of the C5protein.

Competition of each biotinylated anti-human C5 antibody with eachunlabelled anti-human C5 antibody in 100-fold excess was tested in anELISA (capture mode). It was compared with the highest signal of eachantibody (biotinylated Fab without competition).

Human C5 was captured via a polyclonal anti-human C5 IgG (USBiological), which was coated previously o/n at 4° C. on a 384 wellblack Maxisorp plates. Next day the plate was washed twice with PBST andblocked for 2 h with 3% BSA-PBST. After washing 3× with PBST, 20 μlhuman C5 was added and incubated 2 h ar RT. The plate was washed 3× withPBST before adding the Fabs.

20 μl unlabelled Fab (200 μg/ml or 400 μg/ml) (100-fold excess) wasadded to the wells of a Maxisorp plate and subsequently 20 ng/ml or 40ng/ml of biotinylated Fab. The biotinylated and unlabelled Fabs wereincubated for 1 h at RT. The plate was washed 3× with PBST and Strep-APZymax Streptavidin-Alkaline Phosphatase, ZYMED, Code: 43-8322, Lot:50799648 was added for detection of the biotinylated Fab binding via C5to the plates. AttoPhos substrate (Roche) was added to the plates andFluorescence was read after 5-10 min.

Parental Fabs

C5 was captured (via a polyclonal antibody) and unlabelled FabY wasapplied in excess to biotinylated FabX. Binding of biotinylated FabX tohuman C5 was detected. Six groups of Fabs could be defined: Group 1:MOR06952, 6961; Group 2: MOR06525, 6756, 6757, 6763; Group 3: MOR07087;Group 4: MOR06764, 6776, 7081; Group 5: MOR07089; Group 6: MOR07082,7083, 7084, 7086, 7088, 7090, 7091, 7092, 7093, 7095.

The Fabs were also divided into different epitope binding groups using adifferent method: FabX was immobilized, then FabY pre-incubated withbiotinylated C5 was added. Following groups of Fabs could be defined:Group 1: MOR06952, 6961; Group 2: MOR06525, 6757, 7083; Group 3:MOR07087; Group 4: MOR06763; Group 5: MOR07081; Group 6: MOR07082, 7083,7084, 7086, 7088, 7091, 7092, 7093 (7089 competes with 7084). Theconclusion was drawn that using two different methods, similar resultscould be obtained.

Matured Fabs

In order to complete Fab characterization competition of biotinylatedFab with unlabelled Fab (applied in 100-fold excess) was measured insolution ELISA. Results were compared with the highest signal(biotinylated Fab without competition).

As shown in FIG. 19, biotinylated Fabs compete with identical unlabelledFabs and all Fabs compete for binding to the same or overlappingepitope. These results correlate with epitope binning data for theparental Fabs.

Example 13 Screening of C5 Alpha versus Beta Chain Binders andCompetition Assays

Two ELISA experiments and hemolytic assays were performed to test if aFab was an alpha or beta chain binder as described below.

In the first experiment, Fab was coated on a plate and purified C5 orsupernatant from chimeric C5 preparation (human alpha, mouse beta chain)was added. As a next step 5G1.1 was applied and detection was done viaan anti-human IgG.

In a second experiment, 5G1.1 was coated on a plate, purified C5 orsupernatant from chimeric C5 preparation (human alpha, mouse beta chain)was added, then Fab, which was detected with an anti-Myc antibody.

Reference IgG 5G1.1 recognizes the alpha chain and was used to determineif the MorphSys generated Fabs compete with 5G1.1 for binding. In thehemolytic assays supernatant from chimeric C5 preparation was added tohuman C5-depleted serum and Fabs were tested for inhibition ofhemolysis.

Parental Fabs

FIG. 20 shows the results of an ELISA experiment where the Fabs werecoated on a plate, C5 or supernatant of a chimeric C5 preparation (humanalpha chain and mouse beta chain) was added, then 5G1.1. FIG. 21 showsthe results of an ELISA experiment where purified C5 and supernatantfrom chimeric C5 were captured via 5G1.1.

MOR06525, 6756, 6763 were beta chain binders (bind to C5 but notchimeric C5). Most MOR070XX Fabs (derived from solution pannings) arealpha chain binders (bind to C5 and chimeric C5). MOR06952 and 6961compete with 5G1.1 so they are negative for both C5 and chimeric C5 and,thus, are most likely alpha chain binders as 5G1.1. MOR06757 behaveslike MOR06952 and 6961, i.e. it likely is an alpha chain binder.However, MOR06757 does not inhibit hemolysis of chimeric C5 supernatantspiked into C5-depleted serum, while all the other alpha chain bindersdo (see FIG. 22).

In the hemolytic assay supernatant from chimeric C5 prep was added tohuman C5-depleted serum and Fabs were tested for inhibition ofhemolysis. MOR06525, 6756, 6757 and 6763 did not inhibit hemolysis withchimeric C5 and thus, could be beta chain binders. MOR06952, 6961, 7081,7082, 7083, 7084, 7086, 7087, 7088, 7089, 7090, 7091, 7092, 7093, 7094,7095 inhibited hemolysis and thus could be alpha chain binders.

Example 14 Resistance to Proteolysis

To investigate the structural rigidity of Fabs, resistance of Fabs toproteolysis by thermolysin was performed (thermolysin bacterialprotease, Calbiochem). Fab was incubated with thermolysin(Fab:thermolysin=3:1 (w/w), reaction volume of 8 μL) either at 37° C. orat 55° C. (thermolysin activity is optimal at 55° C.). The reaction wasstopped by adding 4 μL of 0.5 M EDTA and 4 μL of 4×LDS sample buffer(Invitrogen) and the stopped samples were run on 4-12% SDS-PAGE atnon-reducing condition. Proteolysis of Fabs was analyzed by monitoringthe disappearance of Fab bands that were visualized by Coomassiestaining.

Parental Fabs

Parental Fabs were tested for resistance to thermolysin proteolysis at37° C. and 55° C. Fab from a humanized IL-1β antibody was used ascontrol. Most tested Fabs were resistant to degradation by thermolysisat 37° C. up to 90 min. To further differentiate the structural rigidityof Fabs, proteolysis was performed at higher temperature of 55° C. Manyof the Fabs tested were quickly degraded at 55° C. (>90% Fab wasdegraded within 30 min), while some Fabs were still resistant toproteolysis after 90 min (e.g., 7094). The resistant Fabs were suggestedto have a more rigid structure such that they might show better in vivopharmacokinetic properties. Results of these experiments are shown inthe FIG. 23 and FIG. 24.

Matured Fabs

Fabs with the highest potency in hemolytic assays were tested forsensitivity to thermolysin at 37° C. and 55° C. In FIG. 25 and FIG. 26,experiments with derivatives of MOR07086, 7091, 7092 and 7093 are shown.

Results of these tests revealed that derivatives of parentals MOR07091,7092 and 7093 were less sensitive to proteolysis, while MOR07086derivatives were more sensitive to proteolysis.

Example 15 MAC Deposition Assay

As the terminal complement cascade ends up with formation of the MAC,inhibition of MAC formation was a further hint for the antibody abilityto block the complement cascade. The rational was to have an additionalset-up independent of cells and cell behaviour

Zymosan (Sigma), which is an insoluble carbohydrate from the cell wallof yeast, used especially in the immunoassay of the alternative pathway,was coated to activate the Alternative Pathway and IgM (Sigma) wascoated to activate the Classical Pathway for determination of MAC(membrane attack complex) deposition. Fabs were pre-incubated with humanserum (6% for AP, 2% for CP) and added to plate. Percentage (%)inhibition of MAC deposition was calculated for each sample relative tobaseline (EDTA treated human serum) and positive control (human serum),and used to generate the IC₅₀ curve with XLFit.

Parental Fabs

Parental Fabs were used in different concentrations and the maximalinhibition (if applicable also IC50 values) were determined (exampleshown in FIG. 27). Most Fabs completely inhibited MAC depositionindicating blocking of C5 cleavage. Potency and ranking of Fabs weresimilar to data from hemolytic assays.

1. An isolated antibody or an antigen binding fragment thereof thatspecifically binds to a complement C5 protein, wherein said antibody hasan affinity constant (K_(A)) of at least 1×10⁷ M⁻¹, 10⁸ M⁻¹, 10⁹ M⁻¹,10¹⁰ M⁻¹, or 10¹¹ M⁻¹.
 2. The antibody of claim 1 which inhibits thealternative complete pathway as measured by in vitro hemolytic assaywith an IC₅₀ of 20-200 pM.
 3. An antibody, or antigen binding fragmentthereof, that specifically binds to complement C5 protein, and crosscompetes with an antibody described in Table
 1. 4. An antibody, orantigen binding fragment thereof, that binds to the same epitope as anantibody described in Table
 1. 5. The antibody of claim 1, wherein saidantibody is a monoclonal antibody.
 6. The antibody of claim 5, whereinsaid antibody is a human or humanized antibody.
 7. The antibody of claim5, wherein said antibody is a chimeric antibody.
 8. The antibody ofclaim 5, wherein said antibody comprises a human heavy chain constantregion and a human light chain constant region.
 9. The antibody of claim5, wherein said antibody is a single chain antibody.
 10. The antibody ofclaim 5, wherein said antibody is a Fab fragment.
 11. The antibody ofclaim 5, wherein said antibody is a scFv.
 12. The antibody of claim 5binds to both human C5 and cynomolgus C5.
 13. The antibody of claim 5 isan IgG isotype.
 14. The antibody of claim 5, wherein said antibodycomprises a framework in which amino acids have been substituted intothe antibody framework from the respective human VH or VL germlinesequences.
 15. An isolated monoclonal antibody, or an antigen bindingfragment thereof, comprising at least one complementarity determining(CDR) sequence having at least 95% sequence identity to SEQ ID NO: 1, 2,3, 4, 5, 6, 17, 18, 19, 20, 21, 22, 33, 34, 35, 36, 37, 38, 49, 50, 61,62, 63, 64, 65, 66, 77, 78, 89, 95, 101, 107, 113, 119, 120, 131, 132,133, 134, 135, 136, 145, 146, 147, 148, 149, 150, 159, 160, 161, 162,163, 164, 173, 174, 175, 176, 177, 178, 195, 196, 197, 198, 199, 200,209, 226, 235, 236, 237, 238, 239, or 240, wherein said antibody bindsto human C5 protein.
 16. An isolated monoclonal antibody, or an antigenbinding fragment thereof, comprising at least one heavy chain CDRsequence that is identical to SEQ ID NO: 1, 2, 3, 17, 18, 19, 33, 34,35, 49, 61, 62, 63, 77, 77, 95, 107, 113, 119, 132, 131, 133, 145, 146,147, 159, 160, 161, 173, 174, 175, 195, 196, 197, 226, 235, 236, or 237,wherein said antibody binds to human C5 protein.
 17. An isolatedmonoclonal antibody, or an antigen binding fragment thereof, comprisingat least one light chain CDR sequence that is identical to SEQ ID NO: 4,5, 6, 20, 21, 22, 36, 37, 38, 50, 64, 65, 66, 78, 89, 101, 120, 134,135, 136, 148, 149, 150, 162, 163, 164, 176, 177, 178, 198, 199, 200,209, 238, 239, or 240, wherein said antibody binds to human C5 protein.18. An isolated monoclonal antibody, or an antigen binding fragmentthereof, comprising a heavy chain CDR 1 selected from the groupconsisting SEQ ID NOs: 1, 17, 33, 61, 131, 145, 159, 173, 195, and 235;a heavy chain CDR2 selected from the group consisting SEQ ID NOs: 2, 18,34, 49, 62, 77, 95, 107, 113, 119, 132, 146, 160, 174, 196, 226, and236; and a heavy chain CDR3 selected from the group consisting SEQ IDNOs: 3, 19, 35, 63, 133, 147, 161, 175, 197, and 237, wherein saidantibody binds to human C5 protein.
 19. An isolated monoclonal antibody,or an antigen binding fragment thereof, comprising a light chain CDR 1selected from the group consisting SEQ ID NOs: 4, 20, 36, 64, 134, 148,162, 176, 198, and 238; a light chain CDR2 selected from the groupconsisting SEQ ID NOs: 5, 21, 37, 65, 135, 149, 163, 177, 199, and 239;and a light chain CDR3 selected from the group consisting SEQ ID NOs: 6,22, 38, 50, 66, 78, 89, 101, 120, 136, 150, 164, 178, 200, 209, and 240,wherein said antibody binds to human C5 protein.
 20. The antibody ofclaim 18, or an antigen binding fragment thereof, further comprising alight chain CDR1 selected from the group consisting SEQ ID NOs: 4, 20,36, 64, 134, 148, 162, 176, 198, and 238; a light chain CDR2 selectedfrom the group consisting SEQ ID NOs: 5, 21, 37, 65, 135, 149, 163, 177,199, and 239; and a light chain CDR3 selected from the group consistingSEQ ID NOs: 6, 22, 38, 50, 66, 78, 89, 101, 120, 136, 150, 164, 178,200, 209, and
 240. 21. An isolated monoclonal antibody, or an antigenbinding fragment thereof, comprising a heavy chain variable regionhaving at least 95% sequence identity to SEQ ID NO: 7, 23, 39, 51, 67,79, 96, 108, 114, 121, 137, 151, 165, 179, 187, 201, 210, 218, 227, 241,253, 257, 273, 277, or 281, wherein said antibody binds to human C5protein.
 22. An isolated monoclonal antibody, or an antigen bindingfragment thereof, comprising a light chain variable region having atleast 95% sequence identity to SEQ ID NO: 8, 24, 40, 52, 68, 80, 90,102, 122, 138, 152, 166, 180, 188, 202, 211, 219, 228, 242, 261, 265,269, 285, and 289, wherein said antibody binds to human C5 protein. 23.The antibody of claim 21, or an antigen binding fragment thereof,further comprising a light chain variable region having at least 95%sequence identity to SEQ ID NO: 8, 24, 40, 52, 68, 80, 90, 102, 122,138, 152, 166, 180, 188, 202, 211, 219, 228, 242, 261, 265, 269, 285,and
 289. 24. An isolated monoclonal antibody, or an antigen bindingfragment thereof, comprising a heavy chain having at least 95% sequenceidentity to SEQ ID NO: 9, 25, 41, 53, 69, 81, 97, 109, 115, 123, 139,153, 167, 181, 189, 203, 212, 220, 229, 243, 249, 254, 258, 274, 278, or282, wherein said antibody binds to human C5 protein.
 25. An isolatedmonoclonal antibody, or an antigen binding fragment thereof, comprisinga light chain having at least 95% sequence identity to SEQ ID NO: 10,26, 42, 54, 70, 82, 91, 103, 124, 140, 154, 168, 182, 190, 204, 213,221, 230, 244, 251, 262, 266, 270, 286, or 290, wherein said antibodybinds to human C5 protein.
 26. The antibody of claim 24, or an antigenbinding fragment thereof, further comprising a light chain having atleast 95% sequence identity to SEQ ID NO: 10, 26, 42, 54, 70, 82, 91,103, 124, 140, 154, 168, 182, 190, 204, 213, 221, 230, 244, 251, 262,266, 270, 286, or
 290. 27. A pharmaceutical composition comprising anantibody of claim 1 and a pharmaceutically acceptable carrier.
 28. Anucleic acid comprising a nucleotide sequence encoding a polypeptidecomprising a heavy chain variable region having at least 95% sequenceidentity to SEQ ID NO: 7, 23, 39, 51, 67, 79, 96, 108, 114, 121, 137,151, 165, 179, 187, 201, 210, 218, 227, 241, 253, 257, 273, 277, or 281.29. A nucleic acid comprising a nucleotide sequence encoding apolypeptide comprising a light chain variable region having at least 95%sequence identity to SEQ ID NO: 8, 24, 40, 52, 68, 80, 90, 102, 122,138, 152, 166, 180, 188, 202, 211, 219, 228, 242, 261, 265, 269, 285,and
 289. 30. A vector comprising the nucleic acid of claim
 28. 31. Anisolated host cell comprising (1) a recombinant DNA segment encoding aheavy chain of the antibody of claim 18, and (2) a second recombinantDNA segment encoding a light chain of the antibody; wherein said DNAsegments are operably linked to a promoter, and are capable of beingexpressed in said host cell.
 32. The host cell of claim 31, wherein theantibody is a human monoclonal antibody.
 33. The host cell of claim 31that is a non-human mammalian cell line.
 34. A method of treating agerelated macular degeneration comprising administering to a subject inneed thereof an effective amount of a composition comprising an antibodyof claim
 1. 35. The method of claim 34, wherein the subject is a human.36. A method of treating a disease comprising administering to a subjectin need thereof an effective amount of a composition comprising anantibody selected from claims 1-26, wherein said disease is asthma,arthritis, autoimmune heart disease, multiple sclerosis, inflammatorybowel disease, ischemia-reperfusion injuries, Barraquer-Simons Syndrome,hemodialysis, systemic lupus, lupus erythematosus, psoriasis, multiplesclerosis, transplantation, Alzheimer's disease, glomerulonephritis, orMPGN II.
 37. The method of claim 36, wherein said disease isglomerulonephritis.
 38. A method of treating paroxysmal nocturnalhemoglobinuria (PNH) comprising administering to a subject in needthereof an effective amount of a composition comprising the antibody ofclaim
 1. 39. A method of ameliorating a symptom associated withextracorporeal circulation comprising administering to a subject in needthereof an effective amount of a composition comprising the antibody ofclaim
 1. 40. A vector comprising the nucleic acid of claim 29.